One of the major questions being pursued in developmental biology today is, """"""""How are specific genes turned on and off at the appropriate times in a developmental sequence?"""""""" The ultimate goal of the research conducted in our laboratory is to study the modes of regulation involved in differentiation in the life cycle of the dimorphic bacterium Caulobacter crescentus. Work carried out in our laboratory so far has resulted in the development of genetic techniques and the construction of a genetic map for C. crescentus. In addition, techniques for the cloning or identification of specific cloned genes have been elucidated. The work contained in this proposal will involve an analysis of the regulation of the expression of the genes involved in motility and chemotaxis. We will determine when in the cell cycle the flagellar genes are expressed and which genes must be functional for the expression of each gene. These studies will help us to understand which genes are the primary genes involved in the cell cycle dependent regulation of flagellar gene expression. Specifically we propose: 1) To determine the time in the cell cycle when cloned fla genes are expressed. 2) To examine the expression of cloned flagellar genes in strains containing various fla mutations. 3) To examine the expression of the flaF and flaG genes in an in vitro expression system. 4) To analyze the phenomenon of the excretion of unassembled flagellin monomers by flaE and flaH mutants. 5) To establish a correlation between the genetic and physical maps of C. crescentus using pulsed current electrophoresis and a collection of Tn5 insertion mutations. 6) To identify additional fla genes using pulsed-current electrophoresis to determine the map locations of previously unmapped fla mutations. 7) To characterize che mutants unable to respond to particular attractants.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM033580-06
Application #
3283487
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1983-09-23
Project End
1990-08-31
Budget Start
1988-09-01
Budget End
1990-08-31
Support Year
6
Fiscal Year
1988
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
Wang, S P; Sharma, P L; Schoenlein, P V et al. (1993) A histidine protein kinase is involved in polar organelle development in Caulobacter crescentus. Proc Natl Acad Sci U S A 90:630-4
Schoenlein, P V; Lui, J; Gallman, L et al. (1992) The Caulobacter crescentus flaFG region regulates synthesis and assembly of flagellin proteins encoded by two genetically unlinked gene clusters. J Bacteriol 174:6046-53
Khambaty, F M; Ely, B (1992) Molecular genetics of the flgI region and its role in flagellum biosynthesis in Caulobacter crescentus. J Bacteriol 174:4101-9
Ely, B; Ely, T W; Gerardot, C J et al. (1990) Circularity of the Caulobacter crescentus chromosome determined by pulsed-field gel electrophoresis. J Bacteriol 172:1262-6
Driks, A; Schoenlein, P V; DeRosier, D J et al. (1990) A Caulobacter gene involved in polar morphogenesis. J Bacteriol 172:2113-23
Schoenlein, P V; Gallman, L S; Ely, B (1989) Organization of the flaFG gene cluster and identification of two additional genes involved in flagellum biogenesis in Caulobacter crescentus. J Bacteriol 171:1544-53
Schoenlein, P V; Ely, B (1989) Characterization of strains containing mutations in the contiguous flaF, flbT, or flbA-flaG transcription unit and identification of a novel fla phenotype in Caulobacter crescentus. J Bacteriol 171:1554-61
Ely, B; Ely, T W (1989) Use of pulsed field gel electrophoresis and transposon mutagenesis to estimate the minimal number of genes required for motility in Caulobacter crescentus. Genetics 123:649-54
Ely, B; Gerardot, C J (1988) Use of pulsed-field-gradient gel electrophoresis to construct a physical map of the Caulobacter crescentus genome. Gene 68:323-33
Bryan, R; Champer, R; Gomes, S et al. (1987) Separation of temporal control and trans-acting modulation of flagellin and chemotaxis genes in Caulobacter. Mol Gen Genet 206:300-6

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