Sporulation of the bacterium Bacillus subtilis is a relatively simple developmental process that involves a highly ordered program of gene expression and morphological change. The goal of this research is to understand how gene expression is coupled to morphological change during Bacillus sporulation to produce the correct pattern of temporal and spatial gene expression. These studies are of broad significance since all organisms that undergo development must regulate gene expression system to uncover fundamental mechanisms involved in developmental gene regulation. Early in sporulation, the daughter chromosome are partitioned to two compartments, and the subsequent pattern of gene expression in each compartment is distinct. Genetic studies suggest that gene expression in the mother cell compartment is somehow coupled to events occurring in the forespore compartment. The properties of two recently discovered regulators of mother cell-specific gene expression provide tow plausible mechanisms for the coupling phenomenon. One of the regulatory proteins is a sigma subunit of RNa polymerase (called sigmak) that may be synthesized as an inactive precursor. Its processing to the active form could be coupled to forespore events. The other regulatory protein (called the 14kDa protein) is a non-sigma transcription factor whose inactivation has been proposed experiments are designed to test these hypotheses and to define the roles of these two proteins in temporal and spatial gene regulation. Specifically, 1) the binding site of the 14kDa protein in a promoter it represses will be mutated to see if the promoter now becomes active earlier and is uncoupled form forespore events, 2) antibody to the 14kDa protein will be prepared and used to determine its amount and location during sporulation, 3) the sink gene (encoding sigmak) will be fused to an inducible promoter and pro-sigmak will be tested for activity. Antibody to pro-sigmak will be prepared to test directly whether sigmak is made as a precursor during sporulation. If pre-sigmak is inactive, a truncated, active sigk gene will be fused to an induced promoter. The ability to produce pro-sigmak or sigmak at will in cells will permit testing of whether pro-sigmak processing is involved in the coupling between compartments.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
1R29GM043585-01
Application #
3468024
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1989-12-01
Project End
1994-11-30
Budget Start
1989-12-01
Budget End
1990-11-30
Support Year
1
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Michigan State University
Department
Type
Schools of Earth Sciences/Natur
DUNS #
193247145
City
East Lansing
State
MI
Country
United States
Zip Code
48824
Chen, Bin; Himes, Paul; Liu, Yu et al. (2014) Structure of bacterial transcription factor SpoIIID and evidence for a novel mode of DNA binding. J Bacteriol 196:2131-42
Konovalova, Anna; Søgaard-Andersen, Lotte; Kroos, Lee (2014) Regulated proteolysis in bacterial development. FEMS Microbiol Rev 38:493-522
Zhou, Ruanbao; Chen, Kangming; Xiang, Xianling et al. (2013) Features of Pro-?K important for cleavage by SpoIVFB, an intramembrane metalloprotease. J Bacteriol 195:2793-806
Kroos, Lee; Akiyama, Yoshinori (2013) Biochemical and structural insights into intramembrane metalloprotease mechanisms. Biochim Biophys Acta 1828:2873-85
Zhang, Yang; Luethy, Paul M; Zhou, Ruanbao et al. (2013) Residues in conserved loops of intramembrane metalloprotease SpoIVFB interact with residues near the cleavage site in pro-?K. J Bacteriol 195:4936-46
Imamura, Daisuke; Kuwana, Ritsuko; Kroos, Lee et al. (2011) Substrate specificity of SpoIIGA, a signal-transducing aspartic protease in Bacilli. J Biochem 149:665-71
Himes, Paul; McBryant, Steven J; Kroos, Lee (2010) Two regions of Bacillus subtilis transcription factor SpoIIID allow a monomer to bind DNA. J Bacteriol 192:1596-606
Zhou, Ruanbao; Cusumano, Christina; Sui, Dexin et al. (2009) Intramembrane proteolytic cleavage of a membrane-tethered transcription factor by a metalloprotease depends on ATP. Proc Natl Acad Sci U S A 106:16174-9
Harry, Kathryn H; Zhou, Ruanbao; Kroos, Lee et al. (2009) Sporulation and enterotoxin (CPE) synthesis are controlled by the sporulation-specific sigma factors SigE and SigK in Clostridium perfringens. J Bacteriol 191:2728-42