Abstract

The long term goal of the proposed research is to gain a detailed understanding of the regulation of SOS and heat-shock networks in Escherichia coli. One portion of the research will be concerned with an analysis of the newly-isolated css mutations which affect proteolysis and the regulation of the heat-shock response. The css gene(s) will be cloned and their gene products identified, the relationship of the css gene(s) to cellular protease activities will be investigated, the mechanism by which css mutations affect the heat-shock response will be determined, and the regulation of the css gene(s) themselves will be explored. Another portion of the research will be devoted to an analysis of aspects of the regulation of the heat-shock response. This work will include the use of operon and gene fusions to isolate new regulatory mutants and a systematic investigation of properties of the dnaK gene product. Finally, we will continue to analyze the regulation and function of a number of genes that are members of the SOS regulatory network. The studies should offer insights into the interactions between these two global regulatory networks and into the role of proteolytic degradation in cellular regulatory processes. The E. coli SOS and heat-shock regulatory networks are useful as models for regulatory networks induced by DNA damage and stress in more complex organisms.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM028988-06
Application #
3276399
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1981-04-01
Project End
1991-03-31
Budget Start
1986-04-01
Budget End
1987-03-31
Support Year
6
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
Massachusetts Institute of Technology
Department
Type
Schools of Arts and Sciences
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139

  Publications

Barthel, T K; Walker, G C (1999) Inferences concerning the ATPase properties of DnaK and other HSP70s are affected by the ADP kinase activity of copurifying nucleoside-diphosphate kinase. J Biol Chem 274:36670-8
Herman, C; Thevenet, D; Bouloc, P et al. (1998) Degradation of carboxy-terminal-tagged cytoplasmic proteins by the Escherichia coli protease HflB (FtsH). Genes Dev 12:1348-55
Zhang, J; Walker, G C (1998) Interactions of peptides with DnaK and C-terminal DnaK fragments studied using fluorescent and radioactive peptides. Arch Biochem Biophys 356:177-86
Zhang, J; Walker, G C (1996) Identification of elements of the peptide binding site of DnaK by peptide cross-linking. J Biol Chem 271:19668-74
Zhang, J; Lee, M H; Walker, G C (1995) P-azidoiodoacetanilide, a new short photocrosslinker that has greater cysteine specificity than p-azidophenacyl bromide and p-azidobromoacetanilide. Biochem Biophys Res Commun 217:1177-84
McCarty, J S; Walker, G C (1994) DnaK mutants defective in ATPase activity are defective in negative regulation of the heat shock response: expression of mutant DnaK proteins results in filamentation. J Bacteriol 176:764-80
Donnelly, C E; Murli, S; Walker, G C (1994) The groE gene products of Escherichia coli are dispensable for mucA+B(+)-dependent UV mutagenesis. Mutat Res 309:225-33
Bukau, B; Reilly, P; McCarty, J et al. (1993) Immunogold localization of the DnaK heat shock protein in Escherichia coli cells. J Gen Microbiol 139:95-9
Donnelly, C E; Walker, G C (1992) Coexpression of UmuD' with UmuC suppresses the UV mutagenesis deficiency of groE mutants. J Bacteriol 174:3133-9
McCarty, J S; Walker, G C (1991) DnaK as a thermometer: threonine-199 is site of autophosphorylation and is critical for ATPase activity. Proc Natl Acad Sci U S A 88:9513-7

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