The goal of this proposal is to determine how cells integrate metabolic pathways into regulatory networks. In Escherichia coli, the gcvTHP (gcv) operon and gcvB gene are part of a complex network. The gcv operon encodes the glycine cleavage (GCV) enzyme complex that provides one-carbon units for cellular methylation reactions. The gcvB gene encodes two small non-translated RNAs that in turn regulate other pathways (e.g., oppA and dppA, encoding the oligopeptide and dipeptide periplasmic binding proteins, respectively). Both gcv and gcvB are activated by the GcvA protein in response to glycine and repressed by the GcvA + GcvR proteins in the absence of glycine. Because expression of gcvB follows a pattern of regulation qualitatively similar to the gcv operon, we hypothesize that the GcvB RNAs play a role in an integrated cellular response to regulate other genes in conjunction with the GCV enzyme system. Using genetics, physiology and biochemistry, we will address two major questions. First, we will examine regulation of the gcvB gene in response to environmental signals and characterize the two RNA transcripts produced with respect to their relative amounts, stabilities and secondary structures. Second, we will determine the scope of the regulatory network and the molecular mechanisms of control by the GcvB RNAs. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM069506-04
Application #
7489840
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Anderson, James J
Project Start
2005-09-01
Project End
2010-02-28
Budget Start
2008-09-01
Budget End
2010-02-28
Support Year
4
Fiscal Year
2008
Total Cost
$272,721
Indirect Cost
Name
University of Iowa
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
062761671
City
Iowa City
State
IA
Country
United States
Zip Code
52242
Pulvermacher, Sarah C; Stauffer, Lorraine T; Stauffer, George V (2009) The small RNA GcvB regulates sstT mRNA expression in Escherichia coli. J Bacteriol 191:238-48
Pulvermacher, Sarah C; Stauffer, Lorraine T; Stauffer, George V (2009) Role of the sRNA GcvB in regulation of cycA in Escherichia coli. Microbiology 155:106-14
Pulvermacher, Sarah C; Stauffer, Lorraine T; Stauffer, George V (2008) The role of the small regulatory RNA GcvB in GcvB/mRNA posttranscriptional regulation of oppA and dppA in Escherichia coli. FEMS Microbiol Lett 281:42-50
McArthur, Sarah D; Pulvermacher, Sarah C; Stauffer, George V (2006) The Yersinia pestis gcvB gene encodes two small regulatory RNA molecules. BMC Microbiol 6:52