Abstract

Site-specific DNA recombination is the central regulatory mechanism in many developmental pathways in prokaryotes and eukaryotes. The chemical mechanism of strand cleavage and strand transfer during site-specific recombination must have overall similarities to mechanisms of general recombination, DNA transposition and RNA splicing. We intend to study the chemical features of the strand breakage and strand joining reactions mediated by the Flp recombinase from Saccharomyces cerevisiae and other related recombinases. We wish to analyze the type of DNA-protein and protein-protein interactions that confer selectivity of phosphoryl transfer and drive strand joining in the recombinant mode. Understanding the chemistry of this reaction may help in the design of drugs that can specifically inhibit this chemistry. Such drugs could be used to prevent undesirable DNA recombination: for example, viral integration. Site- specific recombination also permits targeted integration of genetic information- the basic rationale in gene therapy.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM035654-09
Application #
2177988
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1990-01-01
Project End
1997-12-31
Budget Start
1994-01-01
Budget End
1994-12-31
Support Year
9
Fiscal Year
1994
Total Cost
Indirect Cost

  Institution

Name
University of Texas Austin
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
City
Austin
State
TX
Country
United States
Zip Code
78712

  Publications

Rowley, Paul A; Kachroo, Aashiq H; Ma, Chien-Hui et al. (2010) Electrostatic suppression allows tyrosine site-specific recombination in the absence of a conserved catalytic arginine. J Biol Chem 285:22976-85
Kachroo, Aashiq H; Ma, Chien-Hui; Rowley, Paul A et al. (2010) Restoration of catalytic functions in Cre recombinase mutants by electrostatic compensation between active site and DNA substrate. Nucleic Acids Res 38:6589-601
Ma, Chien-Hui; Rowley, Paul A; Macieszak, Anna et al. (2009) Active site electrostatics protect genome integrity by blocking abortive hydrolysis during DNA recombination. EMBO J 28:1745-56
Ma, Chien-Hui; Kachroo, Aashiq H; Macieszak, Anna et al. (2009) Reactions of Cre with methylphosphonate DNA: similarities and contrasts with Flp and vaccinia topoisomerase. PLoS One 4:e7248
Ma, Chien-Hui; Kwiatek, Agnieszka; Bolusani, Swetha et al. (2007) Unveiling hidden catalytic contributions of the conserved His/Trp-III in tyrosine recombinases: assembly of a novel active site in Flp recombinase harboring alanine at this position. J Mol Biol 368:183-96
Du, Quan; Livshits, Alexei; Kwiatek, Agnieszka et al. (2007) Protein-induced local DNA bends regulate global topology of recombination products. J Mol Biol 368:170-82
Harshey, Rasika M; Jayaram, Makkuni (2006) The mu transpososome through a topological lens. Crit Rev Biochem Mol Biol 41:387-405
Bolusani, Swetha; Ma, Chien-Hui; Paek, Andrew et al. (2006) Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites. Nucleic Acids Res 34:5259-69
Yin, Zhiqi; Jayaram, Makkuni; Pathania, Shailja et al. (2005) The Mu transposase interwraps distant DNA sites within a functional transpososome in the absence of DNA supercoiling. J Biol Chem 280:6149-56
Konieczka, Jay H; Paek, Andrew; Jayaram, Makkuni et al. (2004) Recombination of hybrid target sites by binary combinations of Flp variants: mutations that foster interprotomer collaboration and enlarge substrate tolerance. J Mol Biol 339:365-78

Showing the most recent 10 out of 19 publications