The overall goal of these studies is to determine the mechanisms whereby gene expression is regulated during the process of spore formation in the Gram-positive bacterium Bacillus subtilis. Specifically, we propose to manipulate the exprerssion of genes whose transcription is induced at the onset of sporulation or at intermediate stages of development through the isolation of promoter mutations, regulatory mutations and extragenic suppressor mutations. The effects of these mutations will be analyzed in vivo by employing gene and operon fusions to the lacZ gene of E. coli and in vitro by means of transcription studies with purified forms of E. coli and in vitro by means of transcription studies with purified forms of B. subtilis RNA polymerase holoenzyme. A principal aim of this project is to understand the role of spoO gene products in the initiation phase of sporulation, a problem that will be addressed through studies on the promoters for two genes whose transcription is induced at the onset of sporulation, and through studies on the novel RNA polymerase signa factors that determine their recognition. We willalso examine the regulation of a third gene whose induction is coupled to genetic and morphological events occurring at an intermediate stage of sporulation. In addition, we will develop fusion-generating derivatives of the transposon Tn917 that will create transcription fusions in vivo of B. subtilis genes to the lacZ gene of E. coli or to the luxA and luxB genes of Vibrio fischeri as new tools for the rapid analysis of sporulation genes of many types. Finally, we propose to test genetically the model that RNA polymerase sigma factors are sequence-specific DNA binding proteins that recognize conserved nucleotide sequence signals in the """"""""-35"""""""" and """"""""-10"""""""" regions of their cognate promoters through the isolation of change-of-specificity mutants of the O-37 species of B. subtilis sigma factor. It is anticipated that these studies on cellular differentiation in a highly accessible procaryotic model system may provide significant insights into problems of normal and abnormal differentiation in higher cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM018568-13
Application #
3269316
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1976-02-01
Project End
1990-01-31
Budget Start
1985-02-01
Budget End
1986-01-31
Support Year
13
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Harvard University
Department
Type
Schools of Arts and Sciences
DUNS #
071723621
City
Cambridge
State
MA
Country
United States
Zip Code
DeLoughery, Aaron; Lalanne, Jean-BenoƮt; Losick, Richard et al. (2018) Maturation of polycistronic mRNAs by the endoribonuclease RNase Y and its associated Y-complex in Bacillus subtilis. Proc Natl Acad Sci U S A 115:E5585-E5594
Bradshaw, Niels; Levdikov, Vladimir M; Zimanyi, Christina M et al. (2017) A widespread family of serine/threonine protein phosphatases shares a common regulatory switch with proteasomal proteases. Elife 6:
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Russell, Jonathan R; Cabeen, Matthew T; Wiggins, Paul A et al. (2017) Noise in a phosphorelay drives stochastic entry into sporulation in Bacillus subtilis. EMBO J 36:2856-2869
Wang Erickson, Anna F; Deighan, Padraig; Garcia, Cinthia P et al. (2017) An Amino Acid Substitution in RNA Polymerase That Inhibits the Utilization of an Alternative Sigma Factor. J Bacteriol 199:
Smolentseva, Olga; Gusarov, Ivan; Gautier, Laurent et al. (2017) Mechanism of biofilm-mediated stress resistance and lifespan extension in C. elegans. Sci Rep 7:7137
Wang Erickson, Anna F; Deighan, Padraig; Chen, Shanshan et al. (2017) A novel RNA polymerase-binding protein that interacts with a sigma-factor docking site. Mol Microbiol 105:652-662
DeFrancesco, Alicia S; Masloboeva, Nadezda; Syed, Adnan K et al. (2017) Genome-wide screen for genes involved in eDNA release during biofilm formation by Staphylococcus aureus. Proc Natl Acad Sci U S A 114:E5969-E5978
Flanagan, Kelly A; Comber, Joseph D; Mearls, Elizabeth et al. (2016) A Membrane-Embedded Amino Acid Couples the SpoIIQ Channel Protein to Anti-Sigma Factor Transcriptional Repression during Bacillus subtilis Sporulation. J Bacteriol 198:1451-63
Cabeen, Matthew T; Leiman, Sara A; Losick, Richard (2016) Colony-morphology screening uncovers a role for the Pseudomonas aeruginosa nitrogen-related phosphotransferase system in biofilm formation. Mol Microbiol 99:557-70

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