Abstract

Two type of genetic recombination events will be studied using defined in vitro systems and purified proteins derived from bacteria and yeast. The proteins to be studied are: 1) The FLP protein of the yeast 2 micron plasmid. This protein promotes a site-specific recombination event which results in the inversion of a segment of the 2 micron plasmid DNA relative to the rest of the plasmid. 2) The recA protein of E. coli. This protein plays a central role in homologous or general genetic recombination in bacteria. The long range goal of this project is to elucidate the chemical mechanisms by which these and similar proteins promote genetic recombination. The work will employ the full range of chemical, kinetic, physical, and genetic techniques available for biochemical analysis. Benefits to be derived from this work include an improved understanding of recombination events in eukaryotes, including those involved in the generation of antibody diversity and in the transformation of mammalian cells by a number of DNA tumor viruses, and the development of new technologies in recombinant DNA research.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Modified Research Career Development Award (K04)
Project #
5K04AI000599-03
Application #
3070706
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1984-07-01
Project End
1989-06-30
Budget Start
1986-07-01
Budget End
1987-06-30
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Wisconsin Madison
Department
Type
Earth Sciences/Resources
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715

  Publications

Pugh, B F; Schutte, B C; Cox, M M (1989) Extent of duplex DNA underwinding induced by RecA protein binding in the presence of ATP. J Mol Biol 205:487-92
Lindsley, J E; Cox, M M (1989) Dissociation pathway for recA nucleoprotein filaments formed on linear duplex DNA. J Mol Biol 205:695-711
Bruckner, R C; Cox, M M (1989) The histone-like H protein of Escherichia coli is ribosomal protein S3. Nucleic Acids Res 17:3145-61
Kim, J I; Heuser, J; Cox, M M (1989) Enhanced recA protein binding to Z DNA represents a kinetic perturbation of a general duplex DNA binding pathway. J Biol Chem 264:21848-56
Pugh, B F; Cox, M M (1988) General mechanism for RecA protein binding to duplex DNA. J Mol Biol 203:479-93
Gates, C A; Cox, M M (1988) FLP recombinase is an enzyme. Proc Natl Acad Sci U S A 85:4628-32
Umlauf, S W; Cox, M M (1988) The functional significance of DNA sequence structure in a site-specific genetic recombination reaction. EMBO J 7:1845-52
Senecoff, J F; Rossmeissl, P J; Cox, M M (1988) DNA recognition by the FLP recombinase of the yeast 2 mu plasmid. A mutational analysis of the FLP binding site. J Mol Biol 201:405-21
Pugh, B F; Cox, M M (1988) High salt activation of recA protein ATPase in the absence of DNA. J Biol Chem 263:76-83
Schutte, B C; Cox, M M (1988) Homology-dependent underwinding of duplex DNA in recA protein generated paranemic complexes. Biochemistry 27:7886-94

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