Life Sciences Commons^™

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 …

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 …

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 …

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 …

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 …

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) …

Constructing An Argf- Strain Of Bacillus Subtilis, Allison Faucher, Christine Pybus, Ronald E. Yasbin, Eduardo A. 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 argF gene, which is predicted by bioinformatic analysis to be highly mutable. To achieve this goal, Allison will first construct a non-polar argF genetic …