Abstract

Our preliminary results have established there are at least 2 structural APase genes in B. licheniformis, APase I and APase II, and that the 2 genes are transcribed by different RNA polymerase holoenzymes. We will determine if these are the only 2 genes responsible for the APase species previously localized and isolated from this organism, by using the cloned genes to construct a APase I-APase II double mutant of B. licheniformis. If the double mutant is not APase negative, we will clone the other APase gene(s) and include this cloned gene(s) in all the proposed experiments below. These experiments are designed to examine the hypothesis that the unique distribution of alkaline phosphatase in B. licheniformis ((1) membrane-associated (integral--ecto-protein; peripheral--ecto- and endo-protein) and (2) secreted - primarily in stationary growth) is the result of similar multiple structural genes under different regulatory control. We propose: 1. To determine if there are more than 2 APase structural genes in the B. licheniformis genome and construct B. licheniformis MC14 mutants which retain only 1 functional APase gene. 2. To analyze the transcriptional regulation of each clone APase gene. 3. To determine the DNA sequence of each APase gene and compare the functional domains of the APase proteins as determined from the derived amino acid sequence. 4. To determine the final destination of the APase in APase mutants which retain only 1 APase gene.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM033471-03
Application #
3283233
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1985-01-01
Project End
1988-06-30
Budget Start
1987-01-01
Budget End
1988-06-30
Support Year
3
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
University of Illinois at Chicago
Department
Type
Schools of Arts and Sciences
DUNS #
121911077
City
Chicago
State
IL
Country
United States
Zip Code
60612

  Publications

Kaushal, Bindiya; Paul, Salbi; Hulett, F Marion (2010) Direct regulation of Bacillus subtilis phoPR transcription by transition state regulator ScoC. J Bacteriol 192:3103-13
Puri-Taneja, Ankita; Schau, Matthew; Chen, Yinghua et al. (2007) Regulators of the Bacillus subtilis cydABCD operon: identification of a negative regulator, CcpA, and a positive regulator, ResD. J Bacteriol 189:3348-58
Eldakak, Amr; Hulett, F Marion (2007) Cys303 in the histidine kinase PhoR is crucial for the phosphotransfer reaction in the PhoPR two-component system in Bacillus subtilis. J Bacteriol 189:410-21
Puri-Taneja, Ankita; Paul, Salbi; Chen, Yinghua et al. (2006) CcpA causes repression of the phoPR promoter through a novel transcription start site, P(A6). J Bacteriol 188:1266-78
Abdel-Fattah, Wael R; Chen, Yinghua; Eldakak, Amr et al. (2005) Bacillus subtilis phosphorylated PhoP: direct activation of the E(sigma)A- and repression of the E(sigma)E-responsive phoB-PS+V promoters during pho response. J Bacteriol 187:5166-78
Schau, Matthew; Chen, Yinghua; Hulett, F Marion (2004) Bacillus subtilis YdiH is a direct negative regulator of the cydABCD operon. J Bacteriol 186:4585-95
Paul, Salbi; Birkey, Stephanie; Liu, Wei et al. (2004) Autoinduction of Bacillus subtilis phoPR operon transcription results from enhanced transcription from EsigmaA- and EsigmaE-responsive promoters by phosphorylated PhoP. J Bacteriol 186:4262-75
Chen, Yinghua; Abdel-Fattah, Wael R; Hulett, F Marion (2004) Residues required for Bacillus subtilis PhoP DNA binding or RNA polymerase interaction: alanine scanning of PhoP effector domain transactivation loop and alpha helix 3. J Bacteriol 186:1493-502
Schau, Matthew; Eldakak, Amr; Hulett, F Marion (2004) Terminal oxidases are essential to bypass the requirement for ResD for full Pho induction in Bacillus subtilis. J Bacteriol 186:8424-32
Birck, Catherine; Chen, Yinghua; Hulett, F Marion et al. (2003) The crystal structure of the phosphorylation domain in PhoP reveals a functional tandem association mediated by an asymmetric interface. J Bacteriol 185:254-61

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