Srs2 removes deadly recombination intermediates independently of its interaction with SUMO-modified PCNA.
Saccharomyces cerevisiae Srs2 helicase plays at least two distinct functions. One is to prevent recombinational repair through its recruitment by sumoylated Proliferating Cell Nuclear Antigen (PCNA), evidenced in postreplication-repair deficient cells, and a second one is to eliminate potentially lethal intermediates formed by recombination proteins. Both actions are believed to involve the capacity of Srs2 to displace Rad51 upon translocation on single-stranded DNA (ssDNA), though a role of its helicase activity may be important to remove some toxic recombination structures. Here, we described two new mutants, srs2R1 and srs2R3, that have lost the ability to hinder recombinational repair in postreplication-repair mutants, but are still able to remove toxic recombination structures. Although the mutants present very similar phenotypes, the mutated proteins are differently affected in their biochemical activities. Srs2R1 has lost its capacity to interact with sumoylated PCNA while the biochemical activities of Srs2R3 are attenuated (ATPase, helicase, DNA binding and ability to displace Rad51 from ssDNA). In addition, crossover (CO) frequencies are increased in both mutants. The different roles of Srs2, in relation to its eventual recruitment by sumoylated PCNA, are discussed.
Le Breton
Cyrille
C
CEA-DSV-Institut de Radiobiologie Cellulaire et Moléculaire, UMR217 CNRS/CEA, F-92265 Fontenay aux Roses, France.
Dupaigne
Pauline
P
Robert
Thomas
T
Le Cam
Eric
E
Gangloff
Serge
S
Fabre
Francis
F
Veaute
Xavier
X
D004247:DNA / Q000648:ultrastructure; D004265:DNA Helicases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004260:DNA Repair; D045643:DNA Repair Enzymes; D004268:DNA-Binding Proteins / Q000235:genetics; D017353:Gene Deletion; D009154:Mutation; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D051135:Rad51 Recombinase / Q000648:ultrastructure; D053484:RecQ Helicases / Q000235:genetics; D011995:Recombination, Genetic; D025842:SUMO-1 Protein / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D013489:Suppression, Genetic; D014466:Ultraviolet Rays
D004268:DNA-Binding Proteins; D018809:Proliferating Cell Nuclear Antigen; C063607:RAD18 protein, S cerevisiae; D025842:SUMO-1 Protein; D029701:Saccharomyces cerevisiae Proteins; C061376:SRS2 protein, S cerevisiae; D004247:DNA; C495151:RAD51 protein, S cerevisiae; D051135:Rad51 Recombinase; C075468:RAD54 protein, S cerevisiae; C090472:SGS1 protein, S cerevisiae; D004265:DNA Helicases; D053484:RecQ Helicases; D045643:DNA Repair Enzymes