Myxococcus xanthus is an excellent prokaryotic system for studying the regulation of development. M. xanthus shows a developmental cycle which is similar to the eukaryotic slime molds such as Dictyostelium discoideum. However, there are several distinct advantages to studying development in M. xanthus, since it is a simple gram-negative bacterium. We propose to study morphogenesis and the molecular mechanisms of control of gene expression during development using M. xanthus as a model system. We will continue to work on the on-going projects such as characterization of development-specific sigma factors, expression of development-specific proteins, GTP-binding proteins and msDNAs. We will also attempt to explore several new aspects of M. xanthus including protein kinases and membrane proteins associated with cell motility and morphogenesis. In this proposal, we will pursue the following specific projects: (1) Identification and characterization of development-specific sigma factors. Using the rpoD gene of M. xanthus [the gene for the vegetative sigma factor (sigA)] as a probe, we will continue to clone various development-specific sigma factors and to investigate their roles in differentiation. (2) Protein U and S. In contrast to protein S (an early developmental protein), protein U is a typical late-developmental protein. The gene for protein U and S will be carried out to elucidate how they are secreted and assembled on the spore surface. (3) GTP binding proteins and protein kinases. In addition to various GTP-binding proteins, recently we have found that M. xanthus contains a gene coding for a protein with significant sequence similarity to eukaryotic protein kinases. We will not only characterize this gene and its role in M. xanthus but also further explore to find other genes for protein kinases. (4) Characterization of the major inner membrane protein. We have found that the inner membrane fraction of M. xanthus contains a protein of 130 kd unusually in a large amount. We have recently cloned the gene for this protein and will characterize it for its possible roles in cellular motility and morphogenesis. (5) RecA proteins. We found that M. xanthus contains two recA genes. We will characterize these genes in terms of their functions in the life cycle. (6) Expression and roles of retrons-Mx65 and -Mx162. M. xanthus contains two independent retrons producing msDNA-Mx65 and msDNA-Mx162, respectively. We will further explore their roles and explore their roles and expression during the life cycle.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM026843-13
Application #
3274289
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1987-04-01
Project End
1995-03-31
Budget Start
1992-04-01
Budget End
1993-03-31
Support Year
13
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Medicine & Dentistry of NJ
Department
Type
Schools of Medicine
DUNS #
622146454
City
Piscataway
State
NJ
Country
United States
Zip Code
08854
Lampson, B; Inouye, M; Inouye, S (2001) The msDNAs of bacteria. Prog Nucleic Acid Res Mol Biol 67:65-91
Ueki, T; Inouye, S (2001) SigB, SigC, and SigE from Myxococcus xanthus homologous to sigma32 are not required for heat shock response but for multicellular differentiation. J Mol Microbiol Biotechnol 3:287-93
Udo, H; Lam, C K; Mori, S et al. (2000) Identification of a substrate for Pkn2, a protein Ser/Thr kinase from Myxococcus xanthus by a novel method for substrate identification. J Mol Microbiol Biotechnol 2:557-63
Inouye, S; Jain, R; Ueki, T et al. (2000) A large family of eukaryotic-like protein Ser/Thr kinases of Myxococcus xanthus, a developmental bacterium. Microb Comp Genomics 5:103-20
Yamanaka, K; Inouye, M; Inouye, S (1999) Identification and characterization of five cspA homologous genes from Myxococcus xanthus. Biochim Biophys Acta 1447:357-65
Otani, M; Kozuka, S; Xu, C et al. (1998) Protein W, a spore-specific protein in Myxococcus xanthus, formation of a large electron-dense particle in a spore. Mol Microbiol 30:57-66
Jain, R; Inouye, S (1998) Inhibition of development of Myxococcus xanthus by eukaryotic protein kinase inhibitors. J Bacteriol 180:6544-50
Ueki, T; Inouye, S (1998) A new sigma factor, SigD, essential for stationary phase is also required for multicellular differentiation in Myxococcus xanthus. Genes Cells 3:371-85
Hanlon, W A; Inouye, M; Inouye, S (1997) Pkn9, a Ser/Thr protein kinase involved in the development of Myxococcus xanthus. Mol Microbiol 23:459-71
Udo, H; Inouye, M; Inouye, S (1997) Biochemical characterization of Pkn2, a protein Ser/Thr kinase from Myxococcus xanthus, a Gram-negative developmental bacterium. FEBS Lett 400:188-92

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