Life Sciences Commons^™

The Role Of A Transcription Factor In Regulating Rice Response To Drought Stress, Diana Ha, Liyuan A. Zhang, Jeffery Shen

Undergraduate Research Opportunities Program (UROP)

The current water shortage is a major concern in regard to our global climate change crisis. A decrease in the availability of water will have direct effects on the development of plants. Some crops, such as Oryza sativa, or commonly known as rice, requires an abundant amount of water for adequate growth. With the water shortage crisis, it will become extremely difficult to harvest such crops to meet the world’s food demand. However, many plants have evolved mechanisms for overcoming and tolerating stresses such as drought. My research focuses on studying the proteins involved with these mechanisms. The WRKY superfamily …

Investigating The Origin Of Coprolites From Three Great Basin Caves, Chelsey Vandrisse, Duane P. Moser, David Rhode

Undergraduate Research Opportunities Program (UROP)

The study of coprolites (mummified feces) is a relatively new endeavor, which enables investigations of the health and diet of ancient people and provides some of the oldest evidence to date for the human habitation in North America (2). In this project, 18 coprolites were examined from archeological digs at three Great Basin caves: the Bonneville Estates Rockshelter (UT), Hidden Cave (NV), and Top of the Terrace Rockshelter (UT). The main objectives were: 1) to verify human origin through the presence of mitochondrial DNA (mtDNA) and 2) assuming human origin, characterize intestinal microflora of Native Americans prior to European contact. …

Isolation, Characterization, And Genome Sequence Of The First Representative Of A Novel Class Within The Chloroflexi That Is Abundant In Some U.S. Great Basin Hot Springs And May Play Important Roles In N And C Cycling, Jeremy A. Dodsworth, Senthil K. Murugapiran, Jonathan Gevorkian, James Han, Tanja Woyke, Susan M. Lucas, Sam Pitluck, Len Pennacchio, Lynne Goodwin, Brian P. Hedlund

Undergraduate Research Opportunities Program (UROP)

A thermophilic, facultatively microaerophilic, heterotrophic bacterium, designated strain JAD2, was isolated from sediments of Great Boiling Spring (GBS), an ~80oC, circumneutral hot spring in the Great Basin GB). The strain grew anaerobically on yeast extract or peptone with an optimal growth temperature of 70-75oC. Growth was stimulated by addition of 0.01 atm O2 to the culture vessel headspace, but was inhibited by higher concentrations (0.2 atm). Cells of JAD2 formed non-motile filaments ranging from 10 to >300 μm in length, which typically decreased in length during stationary phase. 16S rRNA gene-targeted pyrotag sequencing and clone library data suggest that close …

Magnetosome Genes In The Gammaproteobacterium Strain Bw-2, Lucero Rivera, Denis Trubitsyn, Dennis A. Bazylinski

Undergraduate Research Opportunities Program (UROP)

Magnetotactic bacteria (MTB) biomineralize intracellular nanometer-sized, magnetic crystals surrounded by a lipid bilayer membrane known as magnetosomes. These crystals, which consist of magnetite (Fe3O4) or greigite (Fe3S4), causes the cell to align along the geomagnetic field lines as they swim, a phenomenon known as magnetotaxis. Strain BW-2 is a magnetite-producing magnetotactic bacterium isolated from Badwater Basin, Death Valley National Park (California) and is one of only two species of MTB that are known to phylogenetically belong to the Gammaproteobacteria class of the Proteobacteria phylum. The biomineralization of magnetite in magnetotactic bacteria is mediated by a series of genes that include …

Combination Of Virb Binding Site Mutations To Evaluate Collective Impact On Icsp Promoter Activity In Shigella Flexneri, Pashtana Usufuzy, Juan C. Duhart, Maria I. Castellanos, Helen Wing

Undergraduate Research Opportunities Program (UROP)

Shigella flexneri is a gram-negative, invasive bacterial pathogen that afflicts the human colonic epithelium, causing shigellosis, an illness triggering severe dysentery. The World Health Organization cites the disease burden of shigellosis near 90 million episodes and 108,000 deaths per year.

The motility and spread of Shigella is modulated by icsP, a virulence gene. The transcription factor VirB positively regulates many virulence genes encoded by the Shigella virulence plasmid. Two distal binding sites of VirB have been shown to regulate the promoter activity of icsP, despite their location of more than 1 kb upstream of the transcription start site. Five VirB …

Dna Secondary Structures And Their Contribution To Mutagenesis In B. Subtilis Stationary Phase Cells, Carmen Vallin, Holly Martin, Christian Ross, Ronald Yasbin, Eduardo Robleto

Undergraduate Research Opportunities Program (UROP)

It is widely known and accepted that the cause of many mutations in cells are generated during the replication process of actively dividing cells, however more recent research has shown that mutations also arise in non growing conditions, a phenomenon known as stationary phase mutagenesis. Much of what is known come from studies in eukaryotic and bacterial models. It has been proposed that in non~growing cells, the process of transcription plays an important role in mutagenesis. We test the hypothesis that DNA secondary structures, formed during transcription, promote mutagenesis. The transcription-generated structures are speculated to be prone to mutations by …

Inactivation Of Spo0a Gene Increases Stationary Phase Mutagenesis In Bacillus Subtilis, Denisse Reyes, Amanda Prisbrey, Holly Martin, Eduardo Robleto

Undergraduate Research Opportunities Program (UROP)

Stationary phase mutagenesis occurs when a population of cells acquires mutations conferring escape from nongrowing or stress conditions. This type of mutations is observed in nutritionally starved cells. Because the mutations occur after the onset of stress and in cells that are in non-replicative conditions, elucidating the underlying mechanisms contributes novel views to the process of evolution and apply to the formation of cancer in human cells and antibiotic resistance in microbial pathogens. Studies have shown that in Bacillus subtilis, the Mfd protein which is a transcription repair coupling factor is necessary for this phenomenon to occur. Here, we investigate …

Evaluating Species Responses To Climate Change Using Ecological Niche Modeling And Genetic Data, Jeanette Perry

Undergraduate Research Opportunities Program (UROP)

The current and projected future warming trends together with degradation of habitats throughout much of the Great Basin and Columbian Plateau represent real threats to many species occupying these regions. If we can determine the impacts of past climatic changes on the distribution of species, we can obtain a better understanding of the future impacts of projected climatic trends on many species in these regions. My results with the Great Basin Pocket Mouse (Perognathus parvus) may be relevant to conservation ecologists and resource managers attempting to protect several Endangered Species Act candidates, such as the pygmy cottontail (Brachylagus idahoensis). I …

Evaluating Snow Microbial Assemblages, Jenny Lam, Jessica K. Guy, Ryan Brock, Matt Oates, Alison E. Murray

Undergraduate Research Opportunities Program (UROP)

Psychrophiles are organisms that grow optimally below 20C (1). The US Great Basin is home to many mountain peaks with an abundance of alpine snow environments perfect for psychrophilic habitation. We analyzed samples from three different locations, Wheeler Peak, Pacific Crest Trail, and Mount Conness, characterizing and comparing the psychrophilic communities at varying depth intervals in the snow. Polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) showed no notable difference in community structure with depth, but there was a distinct difference when comparing different snow environments (i.e. shaded vs. full sun exposure). The chlorophyll concentration decreased as the …

Evaluation Of Virb Binding Site Contribution To The Regulation Of The Icsp Promoter In Shigella Flexneri, Juan C. Duhart, Maria I. Castellanos, Helen J. Wing

Undergraduate Research Opportunities Program (UROP)

Shigella species are gram-negative, rod-shaped bacteria that are closely related to Escherichia coli. Virulent Shigella spp. are intracellular pathogens that invade, replicate and spread through epithelial cells of the lower intestine and cause bacillary dysentery in humans. This disease is characterized by a robust inflammatory response that results in fever, abdominal pain, and bloody diarrhea (3). According to the CDC, approximately 14,000 cases are reported each year in the United States alone. This number however, does not reflect the actual incidence of this disease as many cases go unreported. The molecular pathogenesis of these bacteria lies in the large virulence …

Modifying The Amino Acid Sequence In The Surface-Exposed Loops Of The Omptin Family Of Proteins To Determine Their Effect On Function, Natiera Magnuson, Eun-Hae Kim, Christian Ross, Helen J. Wing

Undergraduate Research Opportunities Program (UROP)

The omptin family of proteins consists of proteases which lie in the outer membrane of some gram-negative, pathogenic bacteria such as Escherichia coli (OmpT), Shigella flexneri (IcsP), Salmonella typhimurium (PgtE), and Yersinia pestis (Pla). These proteases are highly conserved, sharing approximately 50% sequence identity and a β-barrel shape (fig. 1D). The differences in the structure of these four proteins are in the surface-exposed loop region surrounding the active site, but not in the active site itself [4]. These proteases are important for the virulence of many bacteria. For example, OmpT of E. coli cleaves an antimicrobial peptide secreted by epithelial …

Phylogenetic Studies Of Newly Isolated Freshwater Magnetospirilla Using Cbb And Mam Genes, Nathan Viloria, Christopher T. Lefevre, Dennis A. Bazylinski

Undergraduate Research Opportunities Program (UROP)

The phylogeny and general relatedness of prokaryotes is determined by comparisons of the sequences of rRNA genes, most commonly the 16S rRNA gene. Comparisons between other gene sequences have been used for this purpose and some have supported conclusions from 16S rRNA genes while others have not. In this study, 13 new magnetospirilla were phylogenetically characterized using the sequences of the 16S rRNA gene as well as the genes for forms I and II ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) (cbbL and cbbM, respectively) and for two magnetosome membrane proteins unique to magnetotactic bacteria, mamJ and mamK. Polymerase chain reaction (PCR) with degenerate …

Antibiotic Resistance In Bacillus Subtilis As Affected By Transcriptional Derepression And The Stringent Response, Brandon Eisinger, Ronald E. Yasbin, Eduardo A. Robleto

Undergraduate Research Opportunities Program (UROP)

Bacterial cells under conditions of starvation or prolonged non-lethal selective pressures accumulate mutations in highly transcribed genes. This process is part of cellular programs to increase genetic diversity in conditions of stress, also known as stationary phase or stress-induced mutagenesis. This experiment investigated mutation frequencies for antibiotic resistance as affected by the stringent response. The stringent response is a global cellular process that initiates at the cessation of growth and mediates changes in gene expression that repress synthesis of ribosome components. We used Bacillus subtilis strains that differ in RelA proficiency. The relA gene controls the synthesis of (p)ppGpp, the …

The Small Regulatory Rna Ryhb Regulates Icsa Expression In Shigella Flexneri, Nick Egan, Helen J. Wing

Undergraduate Research Opportunities Program (UROP)

Shigella flexneri is a gram negative non-motile, non-spore forming, rod-shaped bacterium responsible for bacillary dysentery in humans. The master regulator, VirF, initiates a cascade of virulence gene activation by acting as a transcription factor for the gene encoding the global regulator, VirB (1). Production of VirB is also negatively regulated by the regulatory small RNA (sRNA), RyhB 2). Regulatory sRNAs are untranslated RNA molecules involved in the regulation of both transcription and translation. RyhB, a 90 nt sRNA, was first identified in E. coli and subsequently found in all Shigella species. In Shigella this sRNA is maximally expressed in response …

Examination Of Germination Receptors Of B. Subtilis And B. Megaterium, Shruti Patel, Alessio Luinetti, Ernesto Abel-Santos, Eduardo Robleto

Undergraduate Research Opportunities Program (UROP)

Many bacterial species including those in the Bacilli group form spores as a mechanism to survive harsh conditions such as extreme temperature, radiation, chemicals, and nutrient starvation. By forming spores, they can remain metabolically dormant for an extended period and revert to their vegetative form when environment becomes favorable. This resumption of metabolism and growth is marked by a process called germination that is triggered by exogenous nutrients such as amino acids, sugars, and nucleotides. The (Ger) germination receptors that are postulated to respond to these germinants, in the case of B. subtilis and B. megaterium, are a complex of …

Impact Of Historical Climate Change On The Genetic Structure Of The Great Basin Pocket Mouse, Perognathus Parvus, Ashwitha Francis, Mallory E. Eckstut, Brett R. Riddle

Undergraduate Research Opportunities Program (UROP)

The dynamics of genetic patterns, such as genetic differentiation within and between populations and the geographic distribution of genetic lineages, are often influenced by historical events (such as climatic changes) that have substantially impacted regional biodiversity (the study of phylogeography; Avise, 20001). The magnitude of genetic differentiation among populations is often increased if suitable habitat is reduced, because there is restricted migration among populations. When migration is restricted, the populations may become genetically differentiated from conspecific populations and, over time, may become independent lineages that follow distinctive evolutionary trajectories (Stebbins, 19523; Nevo & Beiles, 19892; Thomas et al., 19984). In …

Genetically Modifying Arabidopsis Thaliana With A Gene From Drought-Tolerant Xerophyte Larrea Tridentata (Creosote Bush), Norris Lam, Liyuan A. Zhang, Lingkun Gu, Qingxi J. Shen

Undergraduate Research Opportunities Program (UROP)

L. tridentata, or desert creosote bush, is a xerophytic C3 plant native to the American Southwest, and is known to have evolutionarily developed sophisticated cellular mechanisms to deal with periods of intense abiotic stress. Particularly, complex signaling pathways in L. tridentata allow it to survive in periods of severe water deficiency. Through the findings of Zou et al. [5,6], LtWRKY21 synergistically works with abscisic acid (ABA) to transactivate both ABA-inducible HVA1 and HVA22 promoters. In addition, as ABA and gibberellic acid (GA) pathways are known to act antagonistically. Expectantly, the findings of Zou et al. suggest that LtWRKY21 activates ABA …

Stationary Phase Mutagenesis In Bacillus Subtilis: The Interaction Between Transcription And Error-Prone Replication In Conditions Of Stress, Mary Girard, Eduardo Robleto

Undergraduate Research Opportunities Program (UROP)

While under conditions of stress, non-dividing cells may acquire beneficial mutations. This is referred to as stationary phase mutagenesis, or adaptive mutagenesis. Previous research has shown that actively transcribed genes and those under selective pressure are prone to mutations that confer escape from non-dividing conditions. Accordingly, strains lacking transcription factors have shown a drastically lower number of mutations that confer escape while under amino acid starvation than those observed in the wildtype background. Also, error-prone DNA polymerases are known to be active in cells under stress and it has been shown that strains lacking an error-prone DNA polymerase display reduced …

Attempts To Cultivate Bacteria From Deep Subsurface Aquifers And Mountaintop Plant Communities, Eric D. Hughes, J. C. Bruckner, Duane P. Moser

Undergraduate Research Opportunities Program (UROP)

In the late 1990s, the limits of life were pushed even further when microorganisms were discovered thriving 2.5 km below the surface of the Earth in deep South African gold mines. These very simple communities were dominated by a single species of bacteria from within the phylum, Firmicutes. Desulforudis audaxviator remains unique to a sizeable portion of the South African deep subsurface. At depths below 2.5km, it comprises well over 99% of all organisms present, which presents a unique circumstance in which the environment has provided a natural pure culture. From this naturally occurring pure culture, environmental genomics was applied …

The Role Of Rpoe In Stationary Phase Mutagenesis In Bacillus, Turquoise C. Alexander, Eduardo A. Robleto

Undergraduate Research Opportunities Program (UROP)

Stationary phase mutagenesis is a phenomenon whereby random mutations are generated in non-dividing cells. In order to understand how these mutations arise, we use Bacillus subtilis, a gram positive rod-shaped model organism. It is hypothesize that increased transcription promotes stationary phase mutagenesis in this organism. We therefore examined the role of rpoE, a gene that encodes RNA polymerase ! subunit and proposed to influence efficiency of transcription. To this end, we will first generate a strain bearing a deletion in the rpoE gene. In order to determine if this gene is important for mutagenesis, we will examine the accumulation of …

The Role Of Recn In Stationary Phase Mutagenesis In Bacillus Subtilis, Lauren E. Johnson, Katherine R. Ona, Eduardo A. Robleto, Ronald E. Yasbin

Undergraduate Research Opportunities Program (UROP)

Here, we examine mutagenic programs that are independent of growth, such aspects of the evolutionary process are novel and have been implicated in the formation of cancers in animal cells and the acquisition of antibiotic resistance in animal pathogens. Adaptive or stationary phase mutagenesis is a genetic program to in increase diversity in cells under conditions of stress whereby cells escape non-dividing conditions. Previous research has shown that recombination functions are required to generate mutations that promote growth in Escherichia coli cells starved for carbon. This project tests the hypothesis that recombination functions are required for the generation of mutations …

The Characterization Of Two Diverse Magnetotactic Bacteria: Lems And Mms-1, Ulysses C. Pickard, Paul A. Howse, Dennis A. Bazylinski

Undergraduate Research Opportunities Program (UROP)

Magnetotactic bacteria (MTB) are a diverse group of prokaryotes that biomineralize membrane-bound magnetic crystals known as magnetosomes. The magnetosomes are aligned within the cell and consist of either magnetite (Fe3O4) or greigite (Fe3S4). The biomineralization of magnetosomes consists of several processes including: invagination of the cytoplasmic membrane, iron uptake into the cell and then into the magnetosome membrane vesicle, and crystallization of the mineral phase inside the vesicle. Mam genes control magnetosome biomineralization with most of the genes present in an island called a magnetosome island. Many of the mam genes are conserved between different species of MTB. The genes …

Dna Secondary Structures And Their Contribution To Mutagenesis In B. Subtilis Stationary Phase Cells, Carmen Vallin, Katherine Ona, Chris Ross, Ronald E. Yasbin, Eduardo A. Robleto

Undergraduate Research Opportunities Program (UROP)

It is widely known and accepted that the cause of many mutations in cells are generated during the replication process of actively dividing cells, however more recent research has shown that mutations also arise in non growing conditions, a phenomenon known stationary phase mutagenesis. Much of what is known come from studies in eukaryotic and bacterial models. It is proposed that in nongrowing cells, the process of transcription plays an important role in mutagenesis. I will test the hypothesis that secondary structures formed of DNA generated transcription promote mutagenesis. The sequences transcriptiongenerated structures are speculated to be prone to mutations …

Exploring Diversity Of Nitrate Reducing Thermophiles In Nevada Hot Springs, Jenny Lam, Jeremy A. Dodsworth, Brian P. Hedlund

Undergraduate Research Opportunities Program (UROP)

High rates of denitrification have been measured in Nevada geothermal hot springs, but little is known about the thermophiles that contribute to this activity. We hypothesize that heterotrophic bacteria in the genus Thermus are the most important denitrifiers in the springs. Alternatively, other microorganisms including chemolithotrophs may also be important. To test these hypotheses, several different strategies will be used to try to enrich and isolate nitrate-reducing microorganisms. Isolates will be identified by 16S rRNA gene PCR and sequencing. Subsequently, representative isolates will be chosen for nitrate reductase gene (narG) sequencing and for studies on the kinetics of nitrate reduction …

The Regulation Of The Icsp Promoter Of Shigella Flexneri By The Virulence Factor Virb, Maria Castellanos, Dustin Harrison, Helen Wing

Undergraduate Research Opportunities Program (UROP)

Shigella flexneri is a pathogenic bacterium that causes severe dysentery in humans commonly known as shigellosis. Shigella encodes an outer membrane protease called IcsP. The regulation of icsP expression is under direct control of a transcriptional factor called VirB, which controls the expression of many virulence genes in Shigella. Previous work has shown through deletion analysis of the icsP promoter region that sequences as far as 1368 base pairs upstream of the transcription starting site are important for the regulation of the icsP gene by VirB. However, it is still unclear whether VirB activation requires sequences within the icsP promoter …

Construction Of A Thif Genetic Disruption In Bacillus Subtilis, Kathleen Bradley, Christine Pybus, Ronald Yasbin, Eduardo Robleto

Undergraduate Research Opportunities Program (UROP)

The goal of our research is to determine whether the level of transcription of a gene is correlated with the level of mutation in that gene. One factor involved in the mutability of a transcribed gene is the ability of the single stranded DNA to form secondary stem loop structures (SLS), in the wake of the transcription bubble, that contain unpaired mutable bases. We are interested in correlating the levels of mutation with transcription in the thiF gene, which is predicted by bioinformatic analysis to be highly mutable. To achieve this goal, Kathleen will first construct a non-polar thiF genetic …

Investigation Of Gene And Protein Expression Based On Honey Bee (Apis Mellifera) Aging, Flight Experience, And Behavior, Azucena A. Benito, G. E. Mancinelli, A. Ammons, Michelle M. Elekonich

Undergraduate Research Opportunities Program (UROP)

Honeybees undergo a process of adult behavioral development, spending their first 2-3 weeks working inside the constant environment of the hive. At about 3 weeks of age workers leave the hive as foragers who gather pollen and nectar. Previous research found that bees show an enormous decline in immunity as a result of their transition from regular hive jobs to more difficult foraging activities. Foragers can be forced to go back into hive-tasks, thus becoming “reverted nurses” which may also allow a reversal of immunosenescence. Understanding how this happens could prove to be useful because if there is flexibility in …

Expression Of Thor Does Not Increase Desiccation Resistance In Drosophila Melanogaster, Robert L. Kobey, Deborah K. Hoshizaki, Allen G. Gibbs

Undergraduate Research Opportunities Program (UROP)

Using microarray analysis of Drosophila melanogaster, the Gibbs lab has identified several hundred candidate genes that may be involved in desiccation resistance. One of these genes is Thor, an important downstream target of the TOR/insulin signaling pathway. Preliminary results confirm that Thor plays a role in desiccation resistance. Further research will be needed to verify these results and understand the mechanism by which Thor increases desiccation resistance. This research will also serve as a proof-of-principle for testing microarray-derived hypotheses.

A previous microarray analysis found evidence that down-regulation of protein synthesis might be a cellular response to desiccation through the up-regulation …

Genomic Foundations Of Carbon Fixation In Bacteria Living In Hot Springs, Rachel K. Skinner, Brian P. Hedlund, Jeremy A. Dodsworth

Undergraduate Research Opportunities Program (UROP)

Photosynthesis does not occur above 73°C, so organisms living above this temperature must obtain useable carbon by some other mechanism. It is generally assumed that carbon is fixed by thermophiles through the process of chemolithoautotrophy; however, primary production has never been demonstrated to occur in hot springs >73°C. We have shown that two organisms, Thermocrinis and Pyrobaculum, make up more than 90% of the cells in an 80°C Great Basin hot spring, Great Boiling Spring. We hypothesize that these organisms fix carbon in the hot spring via the reverse tricarboxylic acid (rTCA) cycle. To test this hypothesis we will: i) …

Decoding The Protein Interaction Network - An Approach Integrating Biology And Math, Ryan Huang, Lingkun Gu, Qingxi J. Shen

Undergraduate Research Opportunities Program (UROP)

The WRKY super family is known to play a major role during the plant stress response and development. My project focuses on the protein-protein interaction of an Oryzasativa (rice) transcription factor, OsWRKY71 which functions as the repressor of gibberellins signaling pathway. Previous literature revealed that OsWRKY71 can interact with itself or OsWRKY51 to form dimmers by using bimolecular fluorescence complementation (BiFC). To confirm this result, we use yeast two-hybrid system. As our data showed, OsWRKY71 seems to suppress the reporter gene expression of the conventional yeast two-hybrid system, so we use a modified yeast two-hybrid, Mating-based Split Ubiquitin System (MbSUS). …