One of the major questions being pursued in biology today is: """"""""How are specific genes turned on and off at the appropriate times in a developmental sequence?: The aquatic bacterium Caulobacter crescentus provides a simple system for the study of this question since differentiation occurs twice during the normal cell cycle. In each case, the differentiation is characterized by well-defined events at one pole of the cell. The ultimate goal of the research conducted in our laboratory is to elucidate the modes of regulation of the genes involved in this differentiation. The work contained in this proposal will involve in this differentiation. The work contained in this proposal will involve an analysis of the regulation of the gene expression in C. crescentus. We will examine both biosynthetic genes and those which are expressed at specific times in the cell cycle in order to determine how these genes are regulated. During the course of these studies, we will determine how the expression of these genes is tied to the cell cycle. We propose: 1) To characterize mutants with altered polar organelle development (pod) and to determine the role of pod genes in the regulation and timing of flagellar gene expression. 2) To determine the time in the cell cycle when cloned fla and pod genes are expressed using quantitative mRNA analyses, western analyses, and the fusion of reporter genes to fla promoters in order to monitor gene expression. 3) To use oligonucleotide mutagenesis to determine which nucleotides are essential for the biosynthetic and pod gene promoters and for a newly identified class of flagellar gene promoters. 4) To examine the mechanism by which the flbT repressor protein regulates flagellin gene expression by purifying the FlbT protein and analyzing its interaction with flagellin gene promoters.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM034765-08
Application #
3286293
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1985-04-01
Project End
1994-12-31
Budget Start
1993-01-01
Budget End
1993-12-31
Support Year
8
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of South Carolina at Columbia
Department
Type
Schools of Arts and Sciences
DUNS #
111310249
City
Columbia
State
SC
Country
United States
Zip Code
29208
Leclerc, G; Wang, S P; Ely, B (1998) A new class of Caulobacter crescentus flagellar genes. J Bacteriol 180:5010-9
Mullin, D A; Zies, D L; Mullin, A H et al. (1997) Genetic organization and transposition properties of IS511. Mol Gen Genet 254:456-63
Malakooti, J; Ely, B (1995) Principal sigma subunit of the Caulobacter crescentus RNA polymerase. J Bacteriol 177:6854-60
Wang, S P; Kang, P J; Chen, Y P et al. (1995) Synthesis of the Caulobacter ferredoxin protein, FdxA, is cell cycle controlled. J Bacteriol 177:2901-7
Malakooti, J; Wang, S P; Ely, B (1995) A consensus promoter sequence for Caulobacter crescentus genes involved in biosynthetic and housekeeping functions. J Bacteriol 177:4372-6
Wang, S P; Chen, Y P; Ely, B (1995) A ferredoxin, designated FdxP, stimulates p-hydroxybenzoate hydroxylase activity in Caulobacter crescentus. J Bacteriol 177:2908-11
Tarleton, J C; Malakooti, J; Ely, B (1994) Regulation of Caulobacter crescentus ilvBN gene expression. J Bacteriol 176:3765-74
Yun, C; Ely, B; Smit, J (1994) Identification of genes affecting production of the adhesive holdfast of a marine caulobacter. J Bacteriol 176:796-803
Malakooti, J; Ely, B; Matsumura, P (1994) Molecular characterization, nucleotide sequence, and expression of the fliO, fliP, fliQ, and fliR genes of Escherichia coli. J Bacteriol 176:189-97
Ely, B (1992) DNA sequence of the 3' end of the Caulobacter crescentus 16S rRNA gene. Nucleic Acids Res 20:1423

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