title: Analyses of the yeast Rad51 recombinase A265V mutant reveal different in vivo roles of Swi2-like factors.
abstract: The Saccharomyces cerevisiae Swi2-like factors Rad54 and Rdh54 play multifaceted roles in homologous recombination via their DNA translocase activity. Aside from promoting Rad51-mediated DNA strand invasion of a partner chromatid, Rad54 and Rdh54 can remove Rad51 from duplex DNA for intracellular recycling. Although the in vitro properties of the two proteins are similar, differences between the phenotypes of the null allele mutants suggest that they play different roles in vivo. Through the isolation of a novel RAD51 allele encoding a protein with reduced affinity for DNA, we provide evidence that Rad54 and Rdh54 have different in vivo interactions with Rad51. The mutant Rad51 forms a complex on duplex DNA that is more susceptible to dissociation by Rdh54. This Rad51 variant distinguishes the in vivo functions of Rad54 and Rdh54, leading to the conclusion that two translocases remove Rad51 from different substrates in vivo. Additionally, we show that a third Swi2-like factor, Uls1, contributes toward Rad51 clearance from chromatin in the absence of Rad54 and Rdh54, and define a hierarchy of action of the Swi2-like translocases for chromosome damage repair.
authors.affiliation: Molecular Biophysics and Biochemistry, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Chi
authors.forename: YoungHo
authors.initials: Y
authors.lastname: Kwon
authors.forename: Mari-Liis
authors.initials: ML
authors.lastname: Visnapuu
authors.forename: Isabel
authors.initials: I
authors.lastname: Lam
authors.forename: Sergio R
authors.initials: SR
authors.lastname: Santa Maria
authors.forename: Xiuzhong
authors.initials: X
authors.lastname: Zheng
authors.forename: Anastasiya
authors.initials: A
authors.lastname: Epshtein
authors.forename: Eric C
authors.initials: EC
authors.lastname: Greene
authors.forename: Patrick
authors.initials: P
authors.lastname: Sung
authors.forename: Hannah L
authors.initials: HL
authors.lastname: Klein
mesh_terms: D004247:DNA / Q000378:metabolism; D004265:DNA Helicases / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D026942:DNA Topoisomerases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D016153:Genes, Suppressor; D009154:Mutation; D051135:Rad51 Recombinase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D004247:DNA; C495151:RAD51 protein, S cerevisiae; D051135:Rad51 Recombinase; C075468:RAD54 protein, S cerevisiae; C506745:ULS1 protein, S cerevisiae; D004265:DNA Helicases; D026942:DNA Topoisomerases; C541336:RDH54 protein, S cerevisiae; D045643:DNA Repair Enzymes