The long-range goal of this project is to learn how genes are controlled by remotely-bound activator proteins. This is a problem of great significance because most human genes are controlled in this manner. These genes include oncogenes and others controlling the production of other proteins whose functions are critical for the health of the cells and the organism. The discovery of an analogous process in bacteria provides an entry to study aspects of the complex problem in a simple and accessible system. In the simple bacterium E. coli this process requires the participation of the protein sigma 54. The current proposal centers on learning how sigma 54 modifies the bacterial transcription apparatus so that it can be sensitive to remotely bound activator proteins. First the transcription cycle of genes dependent on sigma 54 will be characterized by in vitro studies. Then the mutually interacting domains on sigma 54, activator and RNA polymerase will be identified. Particular attention will be paid to the eukaryotic - like motifs present on sigma 54 including acidic and glutamine-rich regions. Finally the role of each protein and its sub-domains will be analyzed with regard to where it participates in the enhancer-dependent mechanism. This work should serve as a guide to studies of more complex systems and facilitate the emergence of concepts important to transcription in both human cells and micro-organisms.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM035754-13
Application #
2608848
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1985-12-01
Project End
1999-01-31
Budget Start
1997-12-01
Budget End
1999-01-31
Support Year
13
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Huo, Yi-Xin; Rosenthal, Adam Z; Gralla, Jay D (2008) General stress response signalling: unwrapping transcription complexes by DNA relaxation via the sigma38 C-terminal domain. Mol Microbiol 70:369-78
Rosenthal, Adam Z; Kim, Youngbae; Gralla, Jay D (2008) Poising of Escherichia coli RNA polymerase and its release from the sigma 38 C-terminal tail for osmY transcription. J Mol Biol 376:938-49
Rosenthal, Adam Z; Kim, Youngbae; Gralla, Jay D (2008) Regulation of transcription by acetate in Escherichia coli: in vivo and in vitro comparisons. Mol Microbiol 68:907-17
Gralla, Jay D; Huo, Yi-Xin (2008) Remodeling and activation of Escherichia coli RNA polymerase by osmolytes. Biochemistry 47:13189-96
Gralla, Jay D; Vargas, David R (2006) Potassium glutamate as a transcriptional inhibitor during bacterial osmoregulation. EMBO J 25:1515-21
Kim, Youngbae; Lew, Chih M; Gralla, Jay D (2006) Escherichia coli pfs transcription: regulation and proposed roles in autoinducer-2 synthesis and purine excretion. J Bacteriol 188:7457-63
Gralla, Jay D (2005) Escherichia coli ribosomal RNA transcription: regulatory roles for ppGpp, NTPs, architectural proteins and a polymerase-binding protein. Mol Microbiol 55:973-7
Lew, Chih M; Gralla, Jay D (2004) Nucleotide-dependent isomerization of Escherichia coli RNA polymerase. Biochemistry 43:12660-6
Lee, Shun Jin; Gralla, Jay D (2004) Osmo-regulation of bacterial transcription via poised RNA polymerase. Mol Cell 14:153-62
Lee, Shun Jin; Gralla, Jay D (2003) Open complex formation in vitro by sigma38 (rpoS) RNA polymerase: roles for region 2 amino acids. J Mol Biol 329:941-8

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