Showing 151 - 200 of 274 Items

Date: 2021-01-01

Creator: Grace Marie Hambelton

Access: Open access

Baroreceptors are stretch receptors located in the aorta of mammals; in response to increased afterload, they elicit a decrease in heart rate, creating a negative feedback loop that lowers blood pressure. Although lobsters (Homarus americanus) do not have baroreceptors like mammals, closely related land crabs have been shown to have baroreceptor-like responses. Heart contraction is also regulated by the Frank-Starling response, where increasing stretch or preload increases the contractile force of the heart. In addition to these types of biomechanical modulations, lobsters use a central pattern generator, the cardiac ganglion, to maintain synchronicity of the heartbeat. The heart is also controlled by the central nervous system via neuromodulators, such as myosuppressin, which has been shown to increase active force and decrease frequency in isolated lobster hearts. We performed experiments on a lobster heart with the main arteries still intact, and varied the preload by stretching anterior arteries, and the afterload by elevating the dorsal abdominal artery. We added myosuppressin to modulate the cardiac ganglion output and muscle contraction. We found that the baroreceptor-like response is most directly modulated by active force, whereas frequency could be a secondary control. Increasing preload does increase active force, but that does not correlate to a higher cardiac output, which shows that how hard the heart pumps is not what determines how effectively it is pumping. Additionally, we found that myosuppressin has a much stronger effect on frequency than active force, and so with myosuppressin, frequency becomes the main determinant of cardiac output.

Date: 2013-01-01

Creator: Alex H. Williams, Molly A. Kwiatkowski, Adam L. Mortimer, Eve Marder, Mary Lou, Zeeman, Patsy S. Dickinson

Access: Open access

The cardiac ganglion (CG) of Homarus americanus is a central pattern generator that consists of two oscillatory groups of neurons: "small cells" (SCs) and "large cells" (LCs). We have shown that SCs and LCs begin their bursts nearly simultaneously but end their bursts at variable phases. This variability contrasts with many other central pattern generator systems in which phase is well maintained. To determine both the consequences of this variability and how CG phasing is controlled, we modeled the CG as a pair of Morris-Lecar oscillators coupled by electrical and excitatory synapses and constructed a database of 15,000 simulated networks using random parameter sets. These simulations, like our experimental results, displayed variable phase relationships, with the bursts beginning together but ending at variable phases. The model suggests that the variable phasing of the pattern has important implications for the functional role of the excitatory synapses. In networks in which the two oscillators had similar duty cycles, the excitatory coupling functioned to increase cycle frequency. In networks with disparate duty cycles, it functioned to decrease network frequency. Overall, we suggest that the phasing of the CG may vary without compromising appropriate motor output and that this variability may critically determine how the network behaves in response to manipulations. © 2013 the American Physiological Society.

Date: 2007-05-15

Creator: Elizabeth A. Stemmler, Emily A. Bruns, Noah P. Gardner, Patsy S. Dickinson, Andrew E., Christie

Access: Open access

In invertebrates, peptides possessing the carboxy (C)-terminal motif -RXRFamide have been proposed as the homologs of vertebrate neuropeptide Y (NPY). Using matrix assisted laser desorption/ionization mass spectrometry, in combination with sustained off-resonance irradiation collision-induced dissociation and chemical and enzymatic reactions, we have identified the peptide pEGFYSQRYamide from the neuroendocrine pericardial organ (PO) of the crab Pugettia producta. This peptide is likely the same as that previously reported, but misidentified, as PAFYSQRYamide in several earlier reports (e.g. [Li, L., Kelley, W.P., Billimoria, C.P., Christie, A.E., Pulver, S.R., Sweedler, J.V., Marder, E. 2003. Mass spectrometric investigation of the neuropeptide complement and release in the pericardial organs of the crab, Cancer borealis. J. Neurochem. 87, 642-656; Fu, Q., Kutz, K.K., Schmidt, J.J., Hsu, Y.W., Messinger, D.I., Cain, S.D., de la Iglesia, H.O., Christie, A.E., Li, L. 2005. Hormone complement of the Cancer productus sinus gland and pericardial organ: an anatomical and mass spectrometric investigation. J. Comp. Neurol. 493, 607-626.]). The -QRYamide motif contained in pEGFYSQRYamide is identical to that present in many vertebrate members of the NPY superfamily. Mass spectrometric analysis conducted on the POs of several other decapods showed that pEGFYSQRYamide is present in three other brachyurans (Cancer borealis, Cancer irroratus and Cancer productus) as well as in one species from another decapod infraorder (Lithodes maja, an anomuran). Thus, our findings show that at least some invertebrates possess NPY-like peptides in addition to those exhibiting an -RXRFamide C-terminus, and raise the question as to whether the invertebrate -QRYamides are functionally and/or evolutionarily related to the NPY superfamily. © 2007 Elsevier Inc. All rights reserved.

Date: 2021-01-01

Creator: Utku Ferah

Access: Open access

The role of polymorphisms in protein-coding and non-coding regions of the genome during adaptive evolution has been a long-debated subject in evolutionary biology. Although the importance of coding-sequence polymorphisms during evolution has been well-documented, the influence of non-coding regions of the genome on phenotypic diversity and adaptive evolution remains less clear. Enhancers are cis-regulatory elements that dictate gene transcription rates, times, and locations; enhancers are located in noncoding regions and, when active, exhibit an open-chromatin conformation. In the current study, we identified putative enhancers that differ in chromatin conformation among three natural isolates of Drosophila melanogaster from different parts of the world. The genome-wide numbers of enhancers active in some natural isolates—but inactive in others—will provide insight into the amount of raw material available for evolution due to transcriptional regulatory variation.

• Restriction End Date: 2025-06-01

Date: 2022-01-01

Creator: Sara Elizabeth Nelson

Access: Access restricted to the Bowdoin Community

Date: 2022-01-01

Creator: Fiona G Ralph

Access: Open access

Species interactions are important to organisms and to the ecosystems they inhabit. These interactions, sometimes facilitations, can result in increased resiliency for both species. When facilitation occurs, organisms co-assist with physiological and environmental stressors. As anthropogenic impacts become more stressful for modern organisms, these interactions could offer a solution for many species. Ocean acidification has been shown to be detrimental to many calcifying organisms including oysters. More acidic conditions can slow the process of shell calcification, which can slow growth rates. This effect could directly impact the robust oyster farming business in Midcoast Maine. Because of its possible importance to oyster crops, we assessed the potential of Zostera marina, or eelgrass, to ameliorate the stresses of ocean acidification on farmed Eastern Oysters (Crassotrea virginica). Photosynthesizing organisms such as seagrasses have been shown to locally raise pH, which could create growth refugia for calcifying organisms. While eelgrass has the potential to enhance oyster growth rates, its meadows could also be influencing food availability. To better understand these dynamics, we grew C. virginica in two locations in Harpswell, ME. Crassostrea virginica were split into three habitats at each location: seagrass, fringe, and mudflat, and placed on surface or benthic arrays. We found that seagrass presence and depth interacted to increase shell growth rate. Similarly, Z. marina improved condition index of C. virginica. As ocean acidification worsens, oyster farmers might have to turn to mitigation strategies to ensure profit yield from their labors. Zostera marina could be the solution to their future problems.

• Embargo End Date: 2027-05-16

Date: 2024-01-01

Creator: Benjamin Sewell-Grossman

Access: Embargoed

Date: 2024-01-01

Creator: Sade K. McClean

Access: Open access

Arceuthobium pusillum is a hemiparasite that infects select Picea species. The hosts of A. pusillum do not experience the same symptoms of infection. A. pusillum infections are more fatal to P. marinara, and P. glauca. P. rubens, on the other hand, can survive longer with sustained infection. This presents itself as a contemporary issue because P. glauca, one of the parasite’s most vulnerable hosts, was untethered from ecological competition when old growth forests were subjected to large scale anthropogenic disturbances. These disturbances allowed P. glauca to proliferate, with A. pusillum following. A deeper understanding of the host-species specific responses to A. pusillum infection can broaden general knowledge of parasitic growth and development while also potentially inspiring conservation techniques. This study took advantage of the intrinsic differences between host and parasite to visualize infections in P. rubens and P. glauca, highlighting differences in infection outcome. By illuminating lignin and callose within cross sections of infected P. rubens and P. glauca branches, it was revealed that P. rubens forms dense bands of cells around the cortical strands of infection. These bands form more frequently in P. rubens than in P. glauca and are of a significantly larger area in P. rubens than in P. glauca (t(8), p=0.003, p=0.005). The discovery of the exterior bands is novel and exciting, as the bands are possibly made of callose and potentially facilitate P. rubens survival against A. pusillum infection. The foundational discoveries and results of this study should inspire, and warrant, further analysis.

Date: 2017-05-01

Creator: Robert Barron

Access: Open access

Hybrid zones and their dynamics are important in the understanding of the genetic basis of reproductive isolation and speciation. This study seeks to investigate the hybridization dynamics of a Scarus hybrid swarm within the Tropical Eastern Pacific (TEP) that includes four phenotypically distinct species: S. perrico, S. ghobban, S. rubroviolaceus, and S. compressus. Genetic and population structure analyses of four nuclear loci and a mitochondrial locus revealed that one of the four species, S. compressus, was the result of two different hybrid crosses: S. perrico ✕ S. rubroviolaceus and S. perrico ✕ S. ghobban. A NewHybrids model indicated that most of the S. compressus samples were F1 hybrids, but 21% of the S. compressus sample was classified as “parentals” which could also be explained by the presence of either F2 hybrids or backcrosses with S. compressus phenotypes, given the relatively low power of the nuclear data set (4 loci) to resolve complex hybrid genotypes. Significant mito-nuclear discordance in all three non-hybrid species is consistent with an evolutionary effect of backcrossing between F1 hybrids and “pure” species. This study reveals a relative ease of hybridization between parrotfish taxa separated by an estimated 4.5 million years of isolation and opens the door to further studies on the potential effects of gene flow across old species boundaries and perhaps the formation of new species by hybrid speciation in a diverse clade of tropical reef fish. Elucidating the nature of potentially “deep” F2 crosses and backcrosses within the TEP Scarus hybrid system will allow us to better understand the effects of hybridization on evolution and speciation on both a micro- and macro-ecological scale.

Date: 2017-05-01

Creator: Aidan Fisher Coyle

Access: Open access

Hybrid zones provide natural laboratories to study how specific genes, and interactions among genes, may influence fitness. On the east coast of North America, two separate populations of the European green crab (Carcinus maenas) have been introduced in the last two centuries. An early invasion from Southern Europe colonized New England around 1800, and was followed by a second invasion from Northern Europe to Nova Scotia in the early 1980s (Roman 2006). As these populations hybridize, new combinations of genes potentially adapted to different ends of a thermal spectrum are created in a hybrid zone. To test the hypothesis that mitochondrial and nuclear genes have effects on thermal tolerance, I measured response to cold stress in crabs collected from locations between southern Maine and northern Nova Scotia, and then genotyped the mitochondrial CO1 gene and two nuclear SNPs. Three mitochondrial haplotypes, originally from Northern Europe, had a strong effect on the ability of crabs to right themselves at a temperature of 4.5ºC. Crabs carrying these three haplotypes were 20% more likely to right compared to crabs carrying the haplotype from Southern Europe. The two nuclear SNPs, which were derived from transcriptome sequencing and were strong outliers between Northern and Southern European C. maenas populations, had no effect on righting response at low temperature. These results add C. maenas to the short list of ectotherms in which mitochondrial variation affects thermal tolerance, and suggests that natural selection is shaping the structure of the hybrid zone between the northern and southern populations This discovery of linkage between mitochondrial genotype and thermal tolerance also provides potential insight into the patterns of expansion for invasive populations of C. maenas around the world.

Date: 2025-01-01

Creator: Ryan Minje Kang

Access: Access restricted to the Bowdoin Community

Date: 2025-01-01

Creator: Whitt Dodge

Access: Access restricted to the Bowdoin Community

Date: 2025-01-01

Creator: Oscar Koziol Nigam

Access: Access restricted to the Bowdoin Community

Date: 2010-12-02

Creator: William R. Jackman, James J. Yoo, David W. Stock

Access: Open access

Background. The accessibility of the developing zebrafish pharyngeal dentition makes it an advantageous system in which to study many aspects of tooth development from early initiation to late morphogenesis. In mammals, hedgehog signaling is known to be essential for multiple stages of odontogenesis; however, potential roles for the pathway during initiation of tooth development or in later morphogenesis are incompletely understood. Results. We have identified mRNA expression of the hedgehog ligands shha and the receptors ptc1 and ptc2 during zebrafish pharyngeal tooth development. We looked for, but did not detect, tooth germ expression of the other known zebrafish hedgehog ligands shhb, dhh, ihha, or ihhb, suggesting that as in mammals, only Shh participates in zebrafish tooth development. Supporting this idea, we found that morphological and gene expression evidence of tooth initiation is eliminated in shha mutant embryos, and that morpholino antisense oligonucleotide knockdown of shha, but not shhb, function prevents mature tooth formation. Hedgehog pathway inhibition with the antagonist compound cyclopamine affected tooth formation at each stage in which we applied it: arresting development at early stages and disrupting mature tooth morphology when applied later. These results suggest that hedgehog signaling is required continuously during odontogenesis. In contrast, over-expression of shha had no effect on the developing dentition, possibly because shha is normally extensively expressed in the zebrafish pharyngeal region. Conclusion. We have identified previously unknown requirements for hedgehog signaling for early tooth initiation and later morphogenesis. The similarity of our results with data from mouse and other vertebrates suggests that despite gene duplication and changes in the location of where teeth form, the roles of hedgehog signaling in tooth development have been largely conserved during evolution. © 2010 Jackman et al; licensee BioMed Central Ltd.

Date: 2019-06-12

Creator: Yann Gibert, Eric Samarut, Megan K. Ellis, William R. Jackman, Vincent, Laudet

Access: Open access

The diversity of teeth patterns in actinopterygians is impressive with tooth rows in many locations in the oral and pharyngeal regions. The first-formed tooth has been hypothesized to serve as an initiator controlling the formation of the subsequent teeth. In zebrafish, the existence of the first tooth (named 4 V1) is puzzling as its replacement is induced before the opening of the mouth. Functionally, it has been shown that 4 V1 formation requires fibroblast growth factor (FGF) and retinoic acid (RA) signalling. Here, we show that the ablation of 4 V1 prevents the development of the dental row demonstrating its dependency over it. If endogenous levels of FGF and RA are restored after 4 V1 ablation, embryonic dentition starts again by de novo formation of a first tooth, followed by the dental row. Similarly, induction of anterior ectopic teeth induces subsequent tooth formation, demonstrating that the initiator tooth is necessary and sufficient for dental row formation, probably via FGF ligands released by 4 V1 to induce the formation of subsequent teeth. Our results show that by modifying the formation of the initiator tooth it is possible to control the formation of a dental row. This could help to explain the diversity of tooth patterns observed in actinopterygians and more broadly, how diverse traits evolved through molecular fine-tuning.

Date: 2018-01-01

Creator: Caleb Matthew Gordon

Access: Open access

In the zebrafish pharynx, the first three teeth to form, 3V1, 4V1, and 5V1, have distinct adult and embryonic morphologies, suggesting that these teeth may form using different developmental pathways. Previous studies of gene expression profiles and mutant phenotypes in 3V1, 4V1, and 5V1 have identified four genes that might be involved in dissociating these tooth modules: pitx2b, eve1, pbx1a, and pbx1b. To determine how the developmental roles of these four genes differ across 3V1, 4V1, and 5V1, and obtain a better understanding of how these three teeth develop, I performed CRISPR/Cas9– mediated knockouts in each of these genes, or analyzed embryos from a stable transgenic mutant line where available, and observed the resulting tooth germs and mineralized tooth structures via fluorescence and confocal microscopy. Preliminary results implicate pitx2 as being required for tooth mineralization, offer a possible role for pbx1a, pbx1b, and eve1 in distinguishing the developmental pathway of 3V1, and suggest that 3V1 constitutes a distinct developmental module within the early ventral dentition.

Date: 1998-01-01

Creator: Olaf Ellers, Amy S. Johnson, Philip E. Moberg

Access: Open access

Sea urchin skeletons are strengthened by flexible collagenous ligaments that bind together rigid calcite plates at sutures. Whole skeletons without ligaments (removed by bleaching) broke at lower apically applied forces than did intact, fresh skeletons. In addition, in three-point bending tests on excised plate combinations, sutural ligaments strengthened sutures but not plates. The degree of sutural strengthening by ligaments depended on sutural position; in tensile tests, ambital and adapical sutures were strengthened more than adoral sutures. Adapical sutures, which grow fastest, were also the loosest, suggesting that strengthening by ligaments is associated with growth. In fed, growing urchins, sutures overall were looser than in unfed urchins. Looseness was demonstrated visually and by vibration analysis: bleached skeletons of unfed urchins rang at characteristic frequencies, indicating that sound traveled across tightly fitting sutures; skeletons of fed urchins damped vibrations, indicating loss of vibrational energy across looser sutures. Furthermore, bleached skeletons of fed urchins broke at lower apically applied forces than bleached skeletons of unfed urchins, indicating that the sutures of fed urchins had been held together relatively loosely by sutural ligaments. Thus, the apparently rigid dome-like skeleton of urchins sometimes transforms into a flexible, jointed membrane as sutures loosen and become flexible during growth.

Date: 2021-05-01

Creator: Bruce D. Kohorn, Bridgid E. Greed, Gregory Mouille, Stéphane Verger, Susan L., Kohorn

Access: Open access

Angiosperm cell adhesion is dependent on interactions between pectin polysaccharides which make up a significant portion of the plant cell wall. Cell adhesion in Arabidopsis may also be regulated through a pectin-related signaling cascade mediated by a putative O-fucosyltransferase ESMERALDA1 (ESMD1), and the Epidermal Growth Factor (EGF) domains of the pectin binding Wall associated Kinases (WAKs) are a primary candidate substrate for ESMD1 activity. Genetic interactions between WAKs and ESMD1 were examined using a dominant hyperactive allele of WAK2, WAK2cTAP, and a mutant of the putative O-fucosyltransferase ESMD1. WAK2cTAP expression results in a dwarf phenotype and activation of the stress response and reactive oxygen species (ROS) production, while esmd1 is a suppressor of a pectin deficiency induced loss of adhesion. Here we find that esmd1 suppresses the WAK2cTAP dwarf and stress response phenotype, including ROS accumulation and gene expression. Additional analysis suggests that mutations of the potential WAK EGF O-fucosylation site also abate the WAK2cTAP phenotype, yet only evidence for an N-linked but not O-linked sugar addition can be found. Moreover, a WAK locus deletion allele has no effect on the ability of esmd1 to suppress an adhesion deficiency, indicating WAKs and their modification are not a required component of the potential ESMD1 signaling mechanism involved in the control of cell adhesion. The WAK locus deletion does however affect the induction of ROS but not the transcriptional response induced by the elicitors Flagellin, Chitin and oligogalacturonides (OGs).

Date: 2016-01-01

Creator: E. S. Dickinson, A. S. Johnson, O. Ellers, P. S. Dickinson

Access: Open access

Mechanical and neurophysiological anisotropies mediate three-dimensional responses of the heart of Homarus americanus. Although hearts in vivo are loaded multi-axially by pressure, studies of invertebrate cardiac function typically use uniaxial tests. To generate whole-heart length-tension curves, stretch pyramids at constant lengthening and shortening rates were imposed uniaxially and biaxially along longitudinal and transverse axes of the beating whole heart. To determine whether neuropeptides that are known to modulate cardiac activity in H. americanus affect the active or passive components of these length-tension curves, we also performed these tests in the presence of SGRNFLRFamide (SGRN) and GYSNRNYLRFamide (GYS). In uniaxial and biaxial tests, both passive and active forces increased with stretch along both measurement axes. The increase in passive forces was anisotropic, with greater increases along the longitudinal axis. Passive forces showed hysteresis and active forces were higher during lengthening than shortening phases of the stretch pyramid. Active forces at a given length were increased by both neuropeptides. To exert these effects, neuropeptides might have acted indirectly on the muscle via their effects on the cardiac ganglion, directly on the neuromuscular junction, or directly on the muscles. Because increases in response to stretch were also seen in stimulated motor nerve-muscle preparations, at least some of the effects of the peptides are likely peripheral. Taken together, these findings suggest that flexibility in rhythmic cardiac contractions results from the amplified effects of neuropeptides interacting with the length-tension characteristics of the heart.

Date: 2015-05-20

Creator: Jay J. Falk, Hannah M. Ter Hofstede, Patricia L. Jones, Marjorie M. Dixon, Paul A., Faure, Elisabeth K.V. Kalko, Rachel A. Page

Access: Open access

Many predators and parasites eavesdrop on the communication signals of their prey. Eavesdropping is typically studied as dyadic predator-prey species interactions; yet in nature, most predators target multiple prey species and most prey must evade multiple predator species. The impact of predator communities on prey signal evolution is not well understood. Predators could converge in their preferences for conspicuous signal properties, generating competition among predators and natural selection on particular prey signal features. Alternatively, predator species could vary in their preferences for prey signal properties, resulting in sensory-based niche partitioning of prey resources. In the Neotropics, many substrate-gleaning bats use the mate-attraction songs of male katydids to locate them as prey. We studied mechanisms of niche partitioning in four substrate- gleaning bat species and found they are similar in morphology, echolocation signal design and prey-handling ability, but each species preferred different acoustic features of male song in 12 sympatric katydid species. This divergence in predator preference probably contributes to the coexistence of many substrate-gleaning bat species in the Neotropics, and the substantial diversity in the mate-attraction signals of katydids. Our results provide insight into how multiple eavesdropping predator species might influence prey signal evolution through sensory-based niche partitioning.

Date: 1999-04-02

Creator: Shaun Snyders, Bruce D. Kohorn

Access: Open access

The phosphorylation of proteins within the eukaryotic photosynthetic membrane is thought to regulate a number of photosynthetic processes in land plants and algae. Both light quality and intensity influence protein kinase activity via the levels of reductants produced by the thylakoid electron transport chain. We have isolated a family of proteins called TAKs, Arabidopsis thylakoid membrane threonine kinases that phosphorylate the light harvesting complex proteins. TAK activity is enhanced by reductant and is associated with the photosynthetic reaction center II and the cytochrome b6f complex. TAKs are encoded by a gene family that has striking similarity to transforming growth factor β receptors of metazoans. Thus thylakoid protein phosphorylation may be regulated by a cascade of reductant-controlled membrane-bound protein kinases.

Date: 1980-08-25

Creator: Peter M.M. Rae, Bruce D. Kohorn, Robert P. Wade

Access: Open access

Most repeat units of rDNA in Drosophila virilis are interrupted in the 28S rRNA coding region by an intervening sequence about 10 kb in length; uninterrupted repeats have a length of about 11 kb. We have sequenced the coding/intervening sequence junctions and flanking regions in two independent clones of interrupted rDNA, and the corresponding 28S rRNA coding region in a clone of uninterrupted rDNA. The intervening sequence is terminated at both ends by a direct repeat of a fourteen nucleotide sequence that is present once in the corresponding region of an intact gene. This is a phenomenon associated with transposable elements in other eukaryotes and in prokaryotes, and the Drosophila rDNA intervening sequence is discussed in this context. We have compared more than 200 nucleotides of the D. virilis 28S rRNA gene with sequences of homologous regions of rDNA in Tetrahymena pigmentosa (Wild and Sommer, 1980) and Xenopus laevis (Gourse and Gerbi, 1980): There is 93% sequence homology among the diverse species, so that the rDNA region in question (about two-thirds of the way into the 28S rRNA coding sequence) has been very highly conserved in eukaryote evolution. The intervening sequence in T. pigmentosa is at a site 79 nucleotides upstream from the insertion site of the Drosophila intervening sequence. © 1980 IRL Press Limited.

Date: 2001-08-24

Creator: Shaun Snyders, Bruce D. Kohorn

Access: Open access

To survive fluctuations in quality and intensity of light, plants and algae are able to preferentially direct the absorption of light energy to either one of the two photosystems PSI or PSII. This rapid process is referred to as a state transition and has been correlated with the phosphorylation and migration of the light-harvesting complex protein (LHCP) between PSII and PSI. We show here that thylakoid protein kinases (TAKs) are required for state transitions in Arabidopsis. Antisense TAK1 expression leads to a loss of LHCP phosphorylation and a reduction in state transitions. Preferential activation of PSII causes LHCP to accumulate with PSI, and TAK1 mutants disrupt this process. Finally, TAKs also influence the phosphorylation of multiple thylakoid proteins.

Date: 2013-10-23

Creator: Patricia L. Jones, Michael J. Ryan, Victoria Flores, Rachel A. Page

Access: Open access

Animals can use different sources of information when making decisions. Foraging animals often have access to both self-acquired and socially acquired information about prey. The fringe-lipped bat, Trachops cirrhosus, hunts frogs by approaching the calls that frogs produce to attract mates.We examined howthe reliability of self-acquired prey cues affects social learning of novel prey cues. We trained bats to associate an artificial acoustic cue (mobile phone ringtone) with food rewards. Bats were assigned to treatments in which the trained cue was either an unreliable indicator of reward (rewarded 50% of the presentations) or a reliable indicator (rewarded 100% of the presentations), and they were exposed to a conspecific tutor foraging on a reliable (rewarded 100%) novel cue or to the novel cue with no tutor. Bats whose trained cue was unreliable and who had a tutor were significantly more likely to preferentially approach the novel cue when compared with bats whose trained cue was reliable, and to bats that had no tutor. Reliability of self-acquired prey cues therefore affects social learning of novel prey cues by frog-eating bats. Examining when animals use social information to learn about novel prey is key to understanding the social transmission of foraging innovations. © 2013 The Author(s) Published by the Royal Society.

Date: 2009-07-28

Creator: Ruth Griffin, Anne Sustar, Marianne Bonvin, Richard Binari, Alberto, del Valle Rodriguez, Amber M. Hohl, Jack R. Bateman, Christians Villalta, Elleard Heffern, Didier Grunwald, Chris Bakal, Claude Desplan, Gerold Schubiger, C. Ting Wu, Norbert Perrimon

Access: Open access

In Drosophila melanogaster, widely used mitotic recombination-based strategies generate mosaic flies with positive readout for only one daughter cell after division. To differentially label both daughter cells, we developed the twin spot generator (TSG) technique, which through mitotic recombination generates green and red twin spots that are detectable after the first cell division as single cells. We propose wide applications of TSG to lineage and genetic mosaic studies.

Date: 1999-01-01

Creator: Zachary Wills, Jack Bateman, Christopher A. Korey, Allen Comer, David, Van Vactor

Access: Open access

Genetic analysis of growth cone guidance choice points in Drosophila identified neuronal receptor protein tyrosine phosphatases (RPTPs) as key determinants of axon pathfinding behavior. We now demonstrate that the Drosophila Abl tyrosine kinase functions in the intersegmental nerve b (ISNb) motor choice point pathway as an antagonist of the RPTP Dlar. The function of Abl in this pathway is dependent on an intact catalytic domain. We also show that the Abl phosphoprotein substrate Enabled (Ena) is required for choice point navigation. Both Abl and Ena proteins associate with the Dlar cytoplasmic domain and serve as substrates for Dlar in vitro, suggesting that they play a direct role in the Dlar pathway. These data suggest that Dlar, Abl, and Ena define a phosphorylation state-dependent switch that controls growth cone behavior by transmitting signals at the cell surface to the actin cytoskeleton.

Date: 2021-04-01

Creator: Julian R. Garrison, Joshua S. Caplan, Vladimir Douhovnikoff, Thomas J. Mozdzer, Barry A. Logan

Access: Open access

PREMISE: Biological invasions increasingly threaten native biodiversity and ecosystem services. One notable example is the common reed, Phragmites australis, which aggressively invades North American salt marshes. Elevated atmospheric CO2 and nitrogen pollution enhance its growth and facilitate invasion because P. australis responds more strongly to these enrichments than do native species. We investigated how modifications to stomatal features contribute to strong photosynthetic responses to CO2 and nitrogen enrichment in P. australis by evaluating stomatal shifts under experimental conditions and relating them to maximal stomatal conductance (gwmax) and photosynthetic rates. METHODS: Plants were grown in situ in open-top chambers under ambient and elevated atmospheric CO2 (eCO2) and porewater nitrogen (Nenr) in a Chesapeake Bay tidal marsh. We measured light-saturated carbon assimilation rates (Asat) and stomatal characteristics, from which we calculated gwmax and determined whether CO2 and Nenr altered the relationship between gwmax and Asat. RESULTS: eCO2 and Nenr enhanced both gwmax and Asat, but to differing degrees; gwmax was more strongly influenced by Nenr through increases in stomatal density while Asat was more strongly stimulated by eCO2. There was a positive relationship between gwmax and Asat that was not modified by eCO2 or Nenr, individually or in combination. CONCLUSIONS: Changes in stomatal features co-occur with previously described responses of P. australis to eCO2 and Nenr. Complementary responses of stomatal length and density to these global change factors may facilitate greater stomatal conductance and carbon gain, contributing to the invasiveness of the introduced lineage.

Date: 2018-07-01

Creator: Andrew E. Christie, Andy Yu, Micah G. Pascual, Vittoria Roncalli, Matthew C., Cieslak, Amanda N. Warner, Tess J. Lameyer, Meredith E. Stanhope, Patsy S. Dickinson, J. Joe Hull

Access: Open access

Essentially all organisms exhibit recurring patterns of physiology/behavior that oscillate with a period of ~24-h and are synchronized to the solar day. Crustaceans are no exception, with robust circadian rhythms having been documented in many members of this arthropod subphylum. However, little is known about the molecular underpinnings of their circadian rhythmicity. Moreover, the location of the crustacean central clock has not been firmly established, although both the brain and eyestalk ganglia have been hypothesized as loci. The American lobster, Homarus americanus, is known to exhibit multiple circadian rhythms, and immunodetection data suggest that its central clock is located within the eyestalk ganglia rather than in the brain. Here, brain- and eyestalk ganglia-specific transcriptomes were generated and used to assess the presence/absence of transcripts encoding the commonly recognized protein components of arthropod circadian signaling systems in these two regions of the lobster central nervous system. Transcripts encoding putative homologs of the core clock proteins clock, cryptochrome 2, cycle, period and timeless were found in both the brain and eyestalk ganglia assemblies, as were transcripts encoding similar complements of putative clock-associated, clock input pathway and clock output pathway proteins. The presence and identity of transcripts encoding core clock proteins in both regions were confirmed using PCR. These findings suggest that both the brain and eyestalk ganglia possess all of the molecular components needed for the establishment of a circadian signaling system. Whether the brain and eyestalk clocks are independent of one another or represent a single timekeeping system remains to be determined. Interestingly, while most of the proteins deduced from the identified transcripts are shared by both the brain and eyestalk ganglia, assembly-specific isoforms were also identified, e.g., several period variants, suggesting the possibility of region-specific variation in clock function, especially if the brain and eyestalk clocks represent independent oscillators.

Date: 2009-04-15

Creator: Patsy S. Dickinson, Teerawat Wiwatpanit, Emily R. Gabranski, Rachel J. Ackerman, Jake S., Stevens, Christopher R. Cashman, Elizabeth A. Stemmler, Andrew E. Christie

Access: Open access

The allatostatins comprise three structurally distinct peptide families that regulate juvenile hormone production by the insect corpora allata. A-type family members contain the C-terminal motif -YXFGLamide and have been found in species from numerous arthropod taxa. Members of the B-type family exhibit a -WX6Wamide C-terminus and, like the A-type peptides, appear to be broadly conserved within the Arthropoda. By contrast, members of the C-type family, typified by the unblocked C-terminus -PISCF, a pyroglutamine blocked N-terminus, and a disulfide bridge between two internal Cys residues, have only been found in holometabolous insects, i.e. lepidopterans and dipterans. Here, using transcriptomics, we have identified SYWKQCAFNAVSCFamide (disulfide bridging predicted between the two Cys residues), a known honeybee and water flea C-typelike peptide, from the American lobster Homarus americanus (infraorder Astacidea). Using matrix assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS), a mass corresponding to that of SYWKQCAFNAVSCFamide was detected in the H. americanus brain, supporting the existence of this peptide and its theorized structure. Furthermore, SYWKQCAFNAVSCFamide was detected by MALDI-FTMS in neural tissues from five additional astacideans as well as 19 members of four other decapod infraorders (i.e. Achelata, Anomura, Brachyura and Thalassinidea), suggesting that it is a broadly conserved decapod peptide. In H. americanus, SYWKQCAFNAVSCFamide is capable of modulating the output of both the pyloric circuit of the stomatogastric nervous system and the heart. This is the first demonstration of bioactivity for this peptide in any species.

Date: 2018-07-01

Creator: Harrison P. Fisher, Micah G. Pascual, Sylvia I. Jimenez, David A. Michaelson, Colby T., Joncas, Eleanor D. Quenzer, Andrew E. Christie, Hadley W. Horch

Access: Open access

The auditory system of the cricket, Gryllus bimaculatus, demonstrates an unusual amount of anatomical plasticity in response to injury, even in adults. Unilateral removal of the ear causes deafferented auditory neurons in the prothoracic ganglion to sprout dendrites across the midline, a boundary they typically respect, and become synaptically connected to the auditory afferents of the contralateral ear. The molecular basis of this sprouting and novel synaptogenesis in the adult is not understood. We hypothesize that well-conserved developmental guidance cues may recapitulate their guidance functions in the adult in order to facilitate this compensatory growth. As a first step in testing this hypothesis, we have generated a de novo assembly of a prothoracic ganglion transcriptome derived from control and deafferented adult individuals. We have mined this transcriptome for orthologues of guidance molecules from four well-conserved signaling families: Slit, Netrin, Ephrin, and Semaphorin. Here we report that transcripts encoding putative orthologues of most of the candidate developmental ligands and receptors from these signaling families were present in the assembly, indicating expression in the adult G. bimaculatus prothoracic ganglion.

Date: 2019-08-01

Creator: Samantha K. Barry, Taro Nakamura, Yuji Matsuoka, Christoph Straub, Hadley W., Horch, Cassandra G. Extavour

Access: Open access

Altering gene function in a developing organism is central to different kinds of experiments. While tremendously powerful genetic tools have been developed in traditional model systems, it is difficult to manipulate genes or messenger RNA (mRNA) in most other organisms. At the same time, evolutionary and comparative approaches rely on an exploration of gene function in many different species, necessitating the development and adaptation of techniques for manipulating expression outside currently genetically tractable species. This protocol describes a method for injecting reagents into cricket eggs to assay the effects of a given manipulation on embryonic or larval development. Instructions for how to collect and inject eggs with beveled needles are described. This relatively straightforward technique is flexible and potentially adaptable to other insects. One can gather and inject dozens of eggs in a single experiment, and survival rates for buffer-only injections improve with practice and can be as high as 80%. This technique will support several types of experimental approaches including injection of pharmacological agents, in vitro capped mRNA to express genes of interest, double-stranded RNA (dsRNA) to achieve RNA interference, use of clustered regularly interspaced short palindromic repeats (CRISPR) in concert with CRISPR-associated protein 9 (Cas9) reagents for genomic modification, and transposable elements to generate transient or stable transgenic lines.

Date: 2001-01-01

Creator: Patsy S. Dickinson, Jane Hauptman, John Hetling, Anand Mahadevan

Access: Open access

The neuropeptide red pigment concentrating hormone (RPCH), which we have previously shown to activate the cardiac sac motor pattern and lead to a conjoint gastric mill-cardiac sac pattern in the spiny lobster Panulirus, also activates and modulates the pyloric pattern. Like the activity of gastric mill neurons in RPCH, the pattern of activity in the pyloric neurons is considerably more complex than that seen in control saline. This reflects the influence of the cardiac sac motor pattern, and particularly the upstream inferior ventricular (IV) neurons, on many of the pyloric neurons. RPCH intensifies this interaction by increasing the strength of the synaptic connections between the IV neurons and their targets in the stomatogastric ganglion. At the same time, RPCH enhances postinhibitory rebound in the lateral pyloric (LP) neuron. Taken together, these factors largely explain the complex pyloric pattern recorded in RPCH in Panulirus.

Date: 2016-05-01

Creator: Sabine Y Berzins

Access: Open access

Eelgrass (Zostera marina) is a perennial seagrass that provides many vital ecosystem services including stabilizing sediments, maintaining water clarity, and providing complex habitat in the intertidal and shallow subtidal coastline. Historically, Maine supported dense eelgrass beds in shallow waters surrounding islands and along the coastal mainland. However, in 2012, high population densities of European green crabs (Carcinus maenas), which physically disturb and remove eelgrass as they forage, were correlated with widespread eelgrass declines. Over 55% of the area of eelgrass in Casco Bay was lost, mainly between 2012 and 2014. Eelgrass typically grows in low-oxygen sediments that produce a chemically reducing environment. Sulfate-reducing bacteria in these reduced sediments produce hydrogen sulfide, a toxin that can intrude into eelgrass tissues and impair the plants’ ability to photosynthesize. When eelgrass is not present, sulfide can build up in the pore-water. When eelgrass is present, it can oxygenate the sediments through its roots, thereby preventing the intrusion and buildup of toxic hydrogen sulfide. However, if the substrate is de-vegetated, oxygen levels drop as sedimentary organic matter is decomposed, and the accumulation of sulfides to harmful concentrations in the pore-water may make recolonization of eelgrass difficult or perhaps impossible even in the absence of green crabs. In an effort to monitor characteristics of Casco Bay eelgrass beds and determine spatially where eelgrass may be more likely to recover, four Casco Bay sites with varying degrees of vegetation loss were sampled in 2015 for pore-water sulfide concentration, sediment carbon and nitrogen content, and sediment grain size analysis. Measurements of sulfide concentrations showed correlations with the timing of eelgrass loss, such that vegetated sites had low pore-water sulfide concentrations and sites that had been de-vegetated for longer periods of time had high sulfide concentrations. Carbon and nitrogen content in the sediment was higher at de-vegetated sites, likely due to a higher percentage of finer sediments at those locations. Coarser sediments were more highly vegetated than finer sediments, perhaps displaying a preference of green crabs to forage in finer sediments. Catastrophic loss of eelgrass in Casco Bay has likely led to differences in sulfide levels, carbon and nitrogen content in the sediment, and grain size distribution, depending on degree of vegetation. Eelgrass restoration in Casco Bay will likely be limited by high pore-water sulfide concentrations.

• Restriction End Date: 2027-06-01

Date: 2022-01-01

Creator: Yi Peng Wang

Access: Access restricted to the Bowdoin Community

• Embargo End Date: 2026-05-18

Date: 2023-01-01

Creator: Deva K Holliman

Access: Embargoed

Date: 2022-01-01

Creator: Benjamin Ross

Access: Open access

Remote sensing of solar induced chlorophyll fluorescence (SIF) is a valuable tool in understanding the global carbon cycle. While SIF is highly correlated with photosynthesis at the ecosystem scale, the role that remote sensing of SIF can play at smaller scales is still unclear. The goal of my research was to investigate the ability of SIF to detect changes in pigmentation, photosynthesis, and energy partitioning at the grass canopy and leaf level in response to water stress and abscisic acid (ABA) hormone treatments. Both treatments immediately inhibited photosynthesis by limiting gas exchange through stomatal closure, but SIF declined gradually. Recovery of photosynthesis after alleviation of water stress was not reflected in remote measurements of SIF. I found that senescence in the tips of grasses had been driving changes in remote measurements, which affected remote measurements even when measured leaf-level gas exchange in the lower living tissue recovered. This heterogeneous senescence pattern contextualizes the disconnect between SIF and photosynthesis in stressed turfgrass.

Date: 2021-01-01

Creator: Sophia Walton

Access: Open access

The blue mussel Mytilus edulis alters its phenotype in species-specific ways in response to either green crab (Carcinus maenus) or sea star (Asterias sp.) predation. Previous studies have shown that only sea stars induce changes in abductor muscle morphology, while green crabs generally alter the shape and thickness of shells. In the Western Gulf of Maine, Blue mussels collected from wave protected sites with abundant green crab predators were shown to have significantly thicker shells and larger adductor muscles than mussels collected from wave exposed sites with few green crab predators. The phenotypes of mussels originating from wave-protected and high green crab abundance sites increased the handling time by A. forbesi compared to sites with low wave exposure and high green crab abundance. These results contradict the paradigm that shell thickness trades off with abductor morphology, and I propose that a likely candidate for increased energy allocation to these traits is a decrease in reproductive allocation. My results further suggest that the escalating “arms race” between invasive green crabs and blue mussels in the Western Gulf of Maine is leading to changes in the phenotypic response of mussel populations in ways that are likely impacting sea star foraging dynamics.

Date: 2024-01-25

Creator: Heather Bruce, Hadley Wilson Horch

Access: Open access

Visualizing the expression of genes is a fundamental tool in molecular biology. Traditional colorimetric in situ hybridization using long RNA probes has been a staple for visualizing gene expression but has many drawbacks. In situ HCR v3.0, developed by Choi et. al. 2018, offers improvements over traditional in situs in nearly every aspect: probes can simply be ordered rather than painstakingly cloned and transcribed, which also makes them cost-effective; an HCR takes just three days to complete rather than five or more days; HCR is robust and works well for first-time users; and HCR probes can be multiplexed, allowing four to eight genes to be visualized in a single sample. HCR has been used successfully in many arthropods, including insects (Drosophila, Tribolium), crustaceans (Parhyale, Daphnia, Artemia), and chelicerates (Limulus horseshoe crab, Acanthoscurria tarantula). In this demo, you will learn how to design and order HCR probes as well as best practices for experimental design.

Date: 2015-09-01

Creator: Patsy S. Dickinson, Anirudh Sreekrishnan, Molly A. Kwiatkowski, Andrew E. Christie

Access: Open access

Although the crustacean heart is modulated by a large number of peptides and amines, few of these molecules have been localized to the cardiac ganglion itself; most appear to reach the cardiac ganglion only by hormonal routes. Immunohistochemistry in the American lobster Homarus americanus indicates that pyrokinins are present not only in neuroendocrine organs ( pericardial organ and sinus gland), but also in the cardiac ganglion itself, where pyrokinin-positive terminals were found in the pacemaker cell region, as well as surrounding the motor neurons. Surprisingly, the single pyrokinin peptide identified from H. americanus, FSPRLamide, which consists solely of the conserved FXPRLamide residues that characterize pyrokinins, did not alter the activity of the cardiac neuromuscular system. However, a pyrokinin from the shrimp Litopenaeus vannamei [ADFAFNPRLamide, also known as Penaeus vannamei pyrokinin 2 (PevPK2)] increased both the frequency and amplitude of heart contractions when perfused through the isolated whole heart. None of the other crustacean pyrokinins tested (another from L. vannamei and two from the crab Cancer borealis) had any effect on the lobster heart. Similarly, altering the PevPK2 sequence either by truncation or by the substitution of single amino acids resulted in much lower or no activity in all cases; only the conservative substitution of serine for alanine at position 1 resulted in any activity on the heart. Thus, in contrast to other systems (cockroach and crab) in which all tested pyrokinins elicit similar bioactivities, activation of the pyrokinin receptor in the lobster heart appears to be highly isoform specific.

Date: 2015-12-01

Creator: Andrew E. Christie, Megan Chi, Tess J. Lameyer, Micah G. Pascual, Devlin N., Shea, Meredith E. Stanhope, David J. Schulz, Patsy S. Dickinson

Access: Open access

Peptides are the largest and most diverse class of molecules used for neurochemical communication, playing key roles in the control of essentially all aspects of physiology and behavior. The American lobster, Homarus americanus, is a crustacean of commercial and biomedical importance; lobster growth and reproduction are under neuropeptidergic control, and portions of the lobster nervous system serve as models for understanding the general principles underlying rhythmic motor behavior (including peptidergic neuromodulation). While a number of neuropeptides have been identified from H. americanus, and the effects of some have been investigated at the cellular/systems levels, little is currently known about the molecular components of neuropeptidergic signaling in the lobster. Here, a H. americanus neural transcriptome was generated and mined for sequences encoding putative peptide precursors and receptors; 35 precursor- and 41 receptor-encoding transcripts were identified. We predicted 194 distinct neuropeptides from the deduced precursor proteins, including members of the adipokinetic hormone-corazonin-like peptide, allatostatin A, allatostatin C, bursicon, CCHamide, corazonin, crustacean cardioactive peptide, crustacean hyperglycemic hormone (CHH), CHH precursor-related peptide, diuretic hormone 31, diuretic hormone 44, eclosion hormone, FLRFamide, GSEFLamide, insulin-like peptide, intocin, leucokinin, myosuppressin, neuroparsin, neuropeptide F, orcokinin, pigment dispersing hormone, proctolin, pyrokinin, SIFamide, sulfakinin and tachykinin-related peptide families. While some of the predicted peptides are known H. americanus isoforms, most are novel identifications, more than doubling the extant lobster neuropeptidome. The deduced receptor proteins are the first descriptions of H. americanus neuropeptide receptors, and include ones for most of the peptide groups mentioned earlier, as well as those for ecdysistriggering hormone, red pigment concentrating hormone and short neuropeptide F. Multiple receptors were identified for most peptide families. These data represent the most complete description of the molecular underpinnings of peptidergic signaling in H. americanus, and will serve as a foundation for future gene-based studies of neuropeptidergic control in the lobster.

• Restriction End Date: 2028-06-01

Date: 2023-01-01

Creator: Christoph Anders Tatgenhorst

Access: Access restricted to the Bowdoin Community

• Embargo End Date: 2027-05-16

Date: 2024-01-01

Creator: Sarah Conant

Access: Embargoed

Date: 2017-05-01

Creator: Liam Taylor

Access: Open access

Aerial insectivores have been declining across northeastern North America since the end of the 20th century. The mechanisms and demographic patterns of this decline are unclear. On Kent Island, New Brunswick, Canada, an isolated population of Tree Swallows (Tachycineta bicolor) collapsed between 1987 and 2010. To explore how demographic rates (i.e., survival, reproduction, and immigration) drove the population dynamics of these northeastern aerial insectivores, we combined productivity, population survey, and capture-recapture data in an integrated population model analysis. Neither consistently low juvenile survival rates, adult survival rates, nor clutch size were correlated with population growth rate across years. Alternatively, male and female immigration, hatching success, and fledging success rates were correlated with population growth rate. Because local hatching and fledging success rates cannot influence a population without local recruitment, we argue that the demography of these Tree Swallows is mainly structured by immigration. Parameter-substitution simulations revealed that overall decline was likely even if the population had avoided the worst years of demographic collapse. Breeding Bird Survey comparisons demonstrated how the Kent Island population represents both a demographic and geographical extreme at the edge of a declining region. These demographic patterns highlight the sensitivity, even to the point of local extinction, of some isolated populations to region-scale patterns in the production of potential immigrants.

Date: 2025-01-01

Creator: Abigail Jane Steinwachs

Access: Permanent restriction

Date: 2025-01-01

Creator: Diego Armando DeSousa

Access: Permanent restriction

Date: 2025-01-01

Creator: Nicholas Peter Everin

Access: Access restricted to the Bowdoin Community

Date: 2014-01-01

Creator: Nalini M Nadkarni, Nathaniel T Wheelwright

Access: Open access

The Monteverde Cloud Forest Reserve has captured the worldwide attention of biologists, conservationists, and ecologists and has been the setting for extensive investigation over the past 40 years. Roughly 40,000 ecotourists visit the Cloud Forest each year, and it is often considered the archetypal high-altitude rain forest. “Monteverde: Ecology and Conservation of a Tropical Cloud Forest”, edited by Nalini Nadkarni and Nathaniel T. Wheelwright (Oxford University Press, 2000 and Bowdoin’s Scholar’s Bookshelf. Book 1 ), features synthetic chapters and specific accounts written by more than 100 biologist and local residents, presenting in a single volume everything known in 2000 about the biological diversity of Monteverde, Costa Rica, and how to protect it. The new short chapters written in 2014 by original contributors, and presented here update and expand that knowledge through 2014.

Date: 2012-06-26

Creator: Brian D. Young, David I. Weiss, Cecilia I. Zurita-Lopez, Kristofor J. Webb, Steven G., Clarke, Anne E. McBride

Access: Open access

We have characterized the posttranslational methylation of Rps2, Rps3, and Rps27a, three small ribosomal subunit proteins in the yeast Saccharomyces cerevisiae, using mass spectrometry and amino acid analysis. We found that Rps2 is substoichiometrically modified at arginine-10 by the Rmt1 methyltransferase. We demonstrated that Rps3 is stoichiometrically modified by ω- monomethylation at arginine-146 by mass spectrometric and site-directed mutagenic analyses. Substitution of alanine for arginine at position 146 is associated with slow cell growth, suggesting that the amino acid identity at this site may influence ribosomal function and/or biogenesis. Analysis of the three-dimensional structure of Rps3 in S. cerevisiae shows that arginine-146 makes contacts with the small subunit rRNA. Screening of deletion mutants encoding potential yeast methyltransferases revealed that the loss of the YOR021C gene results in the absence of methylation of Rps3. We demonstrated that recombinant Yor021c catalyzes ω-monomethylarginine formation when incubated with S-adenosylmethionine and hypomethylated ribosomes prepared from a YOR021C deletion strain. Interestingly, Yor021c belongs to the family of SPOUT methyltransferases that, to date, have only been shown to modify RNA substrates. Our findings suggest a wider role for SPOUT methyltransferases in nature. Finally, we have demonstrated the presence of a stoichiometrically methylated cysteine residue at position 39 of Rps27a in a zinc-cysteine cluster. The discovery of these three novel sites of protein modification within the small ribosomal subunit will now allow for an analysis of their functional roles in translation and possibly other cellular processes. © 2012 American Chemical Society.

Date: 2009-08-05

Creator: Anne E. McBride, Ana K. Conboy, Shanique P. Brown, Chaiyaboot Ariyachet, Kate L., Rutledge

Access: Open access

The discovery of roles for arginine methylation in intracellular transport and mRNA splicing has focused attention on the methylated arginine-glycine (RG)-rich domains found in many eukaryotic RNA-binding proteins. Sequence similarity among these highly repetitive RG domains, combined with interactions between RG-rich proteins, raises the question of whether these regions are general interaction motifs or whether there is specificity within these domains. Using the essential Saccharomyces cerevisiae mRNA-binding protein Npl3 (ScNpl3) as a model system, we first tested the importance of the RG domain for protein function. While Npl3 lacking the RG domain could not support growth of cells lacking Npl3, surprisingly, expression of the RG domain alone supported partial growth of these cells. To address the specificity of this domain, we created chimeric forms of ScNpl3 with RG-rich domains of S. cerevisiae nucleolar proteins, Gar1 and Nop1 (ScGar1, ScNop1), or of the Candida albicans Npl3 ortholog (CaNpl3). Whereas the CaNpl3 RG chimeric protein retained nearly wild-type function in S. cerevisiae, the ScGar1 and ScNop1 RG domains significantly reduced Npl3 function and self-association, indicating RG domain specificity. Nuclear localization of Npl3 also requires specific RG sequences, yet heterologous RG domains allow similar modulation of Npl3 transport by arginine methylation.

Date: 2007-07-01

Creator: Anne E. McBride, Cecilia Zurita-Lopez, Anthony Regis, Emily Blum, Ana, Conboy, Shannon Elf, Steven Clarke

Access: Open access

Protein arginine methylation plays a key role in numerous eukaryotic processes, such as protein transport and signal transduction. In Candida albicans, two candidate protein arginine methyltransferases (PRMTs) have been identified from the genome sequencing project. Based on sequence comparison, C. albicans candidate PRMTs display similarity to Saccharomyces cerevisiae Hmt1 and Rmt2. Here we demonstrate functional homology of Hmt1 between C. albicans and S. cerevisiae: CaHmt1 supports growth of S. cerevisiae strains that require Hmt1, and CaHmt1 methylates Npl3, a major Hmt1 substrate, in S. cerevisiae. In C. albicans strains lacking CaHmt1, asymmetric dimethylarginine and ω-monomethylarginine levels are significantly decreased, indicating that Hmt1 is the major C. albicans type I PRMT1. Given the known effects of type I PRMTs on nuclear transport of RNA-binding proteins, we tested whether Hmt1 affects nuclear transport of a putative Npl3 ortholog in C. albicans. CaNpl3 allows partial growth of S. cerevisiae npl3Δ strains, but its arginine-glycine-rich C terminus can fully substitute for that of ScNpl3 and also directs methylation-sensitive association with ScNpl3. Expression of green fluorescent protein-tagged CaNpl3 proteins in C. albicans strains with and without CaHmt1 provides evidence for CaHmt1 facilitating export of CaNpl3 in this fungus. We have also identified the C. albicans Rmt2, a type IV fungus- and plant-specific PRMT, by amino acid analysis of an rmt2Δ/rmt2Δ strain, as well as biochemical evidence for additional cryptic PRMTs. Copyright © 2007, American Society for Microbiology. All Rights Reserved.