Abstract

This project is directed towards elucidating the molecular basis for the expression of phosphate-regulated genes in E. coli. This will be done mainly be using genetic analyses to define the regulation of the alkaline phosphatase (phoA) gene. These studies will include: (1) genetic analysis of the phoB, phoR, and phoM regulator genes and the phoA promoter regulatory region including: (a) deletion analysis of the phoB, R region, (b) the isolation of insertion mutants of and lacZ genetic fusions to the phoB, M, and R genes, (c) construction and use of phoB, M, and R specialized transducing phage and plasmids, (d) mutation analysis of the interaction of the phoB, M, and R gene products and (e) mutation analysis of the phoA promoter-regulatory region; and (2) studies of other phosphate regulated (psi) genes in E. coli including: (a) testing the effect of phosphatase regulator mutants on expression of psi::lac fusions, (b) identification and characterization of the psi+ gene products, and (c) isolation and characterization of new mutants that affect the expression of the psi genes.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM035392-04
Application #
3288041
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1985-01-01
Project End
1989-12-31
Budget Start
1988-01-01
Budget End
1988-12-31
Support Year
4
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
Purdue University
Department
Type
Schools of Arts and Sciences
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907

  Publications

Baker, A S; Ciocci, M J; Metcalf, W W et al. (1998) Insights into the mechanism of catalysis by the P-C bond-cleaving enzyme phosphonoacetaldehyde hydrolase derived from gene sequence analysis and mutagenesis. Biochemistry 37:9305-15
Haldimann, A; Fisher, S L; Daniels, L L et al. (1997) Transcriptional regulation of the Enterococcus faecium BM4147 vancomycin resistance gene cluster by the VanS-VanR two-component regulatory system in Escherichia coli K-12. J Bacteriol 179:5903-13
Haldimann, A; Prahalad, M K; Fisher, S L et al. (1996) Altered recognition mutants of the response regulator PhoB: a new genetic strategy for studying protein-protein interactions. Proc Natl Acad Sci U S A 93:14361-6
Wanner, B L (1996) Signal transduction in the control of phosphate-regulated genes of Escherichia coli. Kidney Int 49:964-7
Metcalf, W W; Jiang, W; Daniels, L L et al. (1996) Conditionally replicative and conjugative plasmids carrying lacZ alpha for cloning, mutagenesis, and allele replacement in bacteria. Plasmid 35:1-13
Fisher, S L; Kim, S K; Wanner, B L et al. (1996) Kinetic comparison of the specificity of the vancomycin resistance VanSfor two response regulators, VanR and PhoB. Biochemistry 35:4732-40
Jiang, W; Metcalf, W W; Lee, K S et al. (1995) Molecular cloning, mapping, and regulation of Pho regulon genes for phosphonate breakdown by the phosphonatase pathway of Salmonella typhimurium LT2. J Bacteriol 177:6411-21
Fisher, S L; Jiang, W; Wanner, B L et al. (1995) Cross-talk between the histidine protein kinase VanS and the response regulator PhoB. Characterization and identification of a VanS domain that inhibits activation of PhoB. J Biol Chem 270:23143-9
Wanner, B L (1994) Molecular genetics of carbon-phosphorus bond cleavage in bacteria. Biodegradation 5:175-84
Metcalf, W W; Wanner, B L (1993) Mutational analysis of an Escherichia coli fourteen-gene operon for phosphonate degradation, using TnphoA' elements. J Bacteriol 175:3430-42

Showing the most recent 10 out of 29 publications