Purification and assays of Saccharomyces cerevisiae homologous recombination proteins

Stephen Van Komen; Margaret Macris; Michael G Sehorn; Patrick Sung

doi:10.1016/S0076-6879(06)08028-1

Purification and assays of Saccharomyces cerevisiae homologous recombination proteins

Methods Enzymol. 2006:408:445-63. doi: 10.1016/S0076-6879(06)08028-1.

Authors

Stephen Van Komen¹, Margaret Macris, Michael G Sehorn, Patrick Sung

Affiliation

¹ Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut, USA.

PMID: 16793386
DOI: 10.1016/S0076-6879(06)08028-1

Abstract

Homologous recombination is an important means of eliminating DNA double strand breaks from chromosomes. The homologous recombination reaction is mediated by the Rad51 recombinase, which requires a number of ancillary factors for maximal efficiency. The development of purification procedures and biochemical assays for yeast Rad51 and other yeast recombination proteins has allowed investigators to begin dissecting the hierarchy of physical and functional interactions among these protein factors that govern the integrity of the homologous recombination machinery. The biochemical studies done with yeast recombination factors have helped formulate conceptual frameworks to guide similar endeavors in other eukaryotes, including humans. Continuing efforts with reconstituted systems that comprise yeast factors will undoubtedly continue to provide insights into the mechanistic intricacy of the homologous recombination machinery.

Publication types

Research Support, U.S. Gov't, P.H.S.

MeSH terms

DNA Damage
DNA Helicases
DNA Repair
DNA Repair Enzymes
DNA, Single-Stranded / metabolism
DNA, Single-Stranded / ultrastructure
Rad51 Recombinase / genetics
Rad51 Recombinase / isolation & purification
Rad51 Recombinase / metabolism
Rad52 DNA Repair and Recombination Protein / genetics
Rad52 DNA Repair and Recombination Protein / isolation & purification
Rad52 DNA Repair and Recombination Protein / metabolism
Recombination, Genetic*
Replication Protein A / genetics
Replication Protein A / isolation & purification
Replication Protein A / metabolism
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae / metabolism
Saccharomyces cerevisiae Proteins* / genetics
Saccharomyces cerevisiae Proteins* / isolation & purification
Saccharomyces cerevisiae Proteins* / metabolism

Substances

DNA, Single-Stranded
Rad52 DNA Repair and Recombination Protein
Replication Protein A
Saccharomyces cerevisiae Proteins
Rad51 Recombinase
RAD54 protein, S cerevisiae
DNA Helicases
DNA Repair Enzymes