We study how the cell controls information flow from genes into proteins. In bacteria, primary control is at the transcriptional level, orchestrated by multiple sigmas (?s). Housekeeping ?s direct transcription from 1000's of promoters;alternative ?s direct transcription from specific genes to cope with environmental change. Our goal is to understand the interplay between ?s and RNA polymerase (RNAP), and the network properties that govern RNAP output. We bring to this work a long history of providing insight into the roles of ?s and a new genomic technology. In the current granting period, we will investigate three fundamental and unsolved issues: (1) The alternative "stress" ?s increase transcription from a limited gene set necessary to cope with the stress;we will determine features of alternative ?s that result in stringent promoter selection. (2) We have been developing new paradigms to predict alternative ?s. Using combinatorial libraries and high throughput approaches, we will critically test the ability of current algorithms to predict promoters of the ?E stress ?, and develop new algorithms that function better. (3) Transcription initiation has been studied largely in isolation. Using our new genomic methodology for high throughput, quantitative determination of genetic interactions, we will determine how RNAP is integrated into the broader workings of the bacterial cell. These benchmark studies will be made available immediately in a public database, thereby moving the entire field forward by identifying new proteins and functional relationships in transcription and new relationships among cellular processes. We perform these studies primarily in E. coli, where a wealth of genetic, biochemical and physiological data facilitate analysis, but our results are directly applicable to all bacteria, including pathogens and environmentally beneficial microbes. Moreover, the questions we are asking and the methods we are developing allow us to directly extend our studies to other bacteria that may be difficult or even impossible to study experimentally. Finally, our studies have important direct applications. The family of circuits we are dissecting may provide building blocks for synthetic circuits with diverse output properties. Additionally, the observation that many RNAP mutants have enhanced growth capabilities under particular conditions or increased resistance to environmental stress indicates that RNAP is a hotspot for adaptive evolution;understanding the mechanism behind these effects could be of use in drug discovery efforts, metabolic engineering, biofuel production and bioremediation.

Public Health Relevance

We study the molecular machineries and circuitries that govern gene expression in bacteria and enable them to proliferate under diverse and challenging environments. This information allows us to manipulate and control bacteria, including pathogens and environmentally beneficial microorganisms. Additionally, our work is of direct relevance to drug discovery, metabolic engineering, biofuel production and bioremediation.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM057755-30
Application #
8300931
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Sledjeski, Darren D
Project Start
1983-01-01
Project End
2014-07-31
Budget Start
2012-08-01
Budget End
2013-07-31
Support Year
30
Fiscal Year
2012
Total Cost
$472,112
Indirect Cost
$161,928
Name
University of California San Francisco
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Rhodius, Virgil A; Segall-Shapiro, Thomas H; Sharon, Brian D et al. (2013) Design of orthogonal genetic switches based on a crosstalk map of ýýs, anti-ýýs, and promoters. Mol Syst Biol 9:702
Rhodius, Virgil A; Mutalik, Vivek K; Gross, Carol A (2012) Predicting the strength of UP-elements and full-length E. coli ýýE promoters. Nucleic Acids Res 40:2907-24
Rhodius, Virgil A; Mutalik, Vivek K (2010) Predicting strength and function for promoters of the Escherichia coli alternative sigma factor, sigmaE. Proc Natl Acad Sci U S A 107:2854-9
Mutalik, Vivek K; Nonaka, Gen; Ades, Sarah E et al. (2009) Promoter strength properties of the complete sigma E regulon of Escherichia coli and Salmonella enterica. J Bacteriol 191:7279-87
Koo, Byoung-Mo; Rhodius, Virgil A; Nonaka, Gen et al. (2009) Reduced capacity of alternative sigmas to melt promoters ensures stringent promoter recognition. Genes Dev 23:2426-36
Rhodius, Virgil A; Wade, Joseph T (2009) Technical considerations in using DNA microarrays to define regulons. Methods 47:63-72
Guisbert, Eric; Rhodius, Virgil A; Ahuja, Nidhi et al. (2007) Hfq modulates the sigmaE-mediated envelope stress response and the sigma32-mediated cytoplasmic stress response in Escherichia coli. J Bacteriol 189:1963-73
Nonaka, Gen; Blankschien, Matthew; Herman, Christophe et al. (2006) Regulon and promoter analysis of the E. coli heat-shock factor, sigma32, reveals a multifaceted cellular response to heat stress. Genes Dev 20:1776-89
Rhodius, Virgil A; Suh, Won Chul; Nonaka, Gen et al. (2006) Conserved and variable functions of the sigmaE stress response in related genomes. PLoS Biol 4:e2
Berghofer-Hochheimer, Yvonne; Lu, Chi Zen; Gross, Carol A (2005) Altering the interaction between sigma70 and RNA polymerase generates complexes with distinct transcription-elongation properties. Proc Natl Acad Sci U S A 102:1157-62

Showing the most recent 10 out of 18 publications