Role for the fission yeast RecQ helicase in DNA repair in G2.
Members of the RecQ helicase subfamily are mutated in several human genomic instability syndromes, such as Bloom, Werner, and Rothmund-Thomson syndromes. We show that Rqh1, the single Schizosaccharomyces pombe homologue, is a 3'-to-5' helicase and exists with Top3 in a high-molecular-weight complex. top3 deletion is inviable, and this is suppressed by concomitant loss of rqh1 helicase activity or loss of recombination functions. This is consistent with RecQ helicases in other systems. By using epistasis analysis of the UV radiation sensitivity and by analyzing the kinetics of Rhp51 (Rad51 homologue), Rqh1, and Top3 focus formation in response to UV in synchronized cells, we identify the first evidence of a function for Rqh1 and Top3 in the repair of UV-induced DNA damage in G(2). Our data provide evidence that Rqh1 functions after Rad51 focus formation during DNA repair. We also identify a function for Rqh1 upstream of recombination in an Rhp18-dependent (Rad18 homologue) pathway. The model that these data allow us to propose helps to reconcile different interpretations of RecQ family helicase function that have arisen between work based on the S. pombe system and models based on studies of Saccharomyces cerevisiae SGS1 suggesting that RecQ helicases act before Rad51.
Laursen
Louise V
LV
Department of Molecular Biology, Aarhus University, DK-8000 Aarhus C, Denmark.
Ampatzidou
Eleni
E
Andersen
Anni H
AH
Murray
Johanne M
JM
D000251:Adenosine Triphosphatases / Q000502:physiology*; D000483:Alleles; D015153:Blotting, Western; D003001:Cloning, Molecular; D004249:DNA Damage; D004265:DNA Helicases / Q000378:metabolism / Q000502:physiology*; D004260:DNA Repair; D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004307:Dose-Response Relationship, Radiation; D016195:G2 Phase; D008954:Models, Biological; D009154:Mutation; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D053484:RecQ Helicases; D011995:Recombination, Genetic; D012568:Schizosaccharomyces; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism; D013347:Subcellular Fractions; D013997:Time Factors; D014466:Ultraviolet Rays
D018076:DNA, Complementary; D029702:Schizosaccharomyces pombe Proteins; D000251:Adenosine Triphosphatases; C506725:RECQL protein, human; D004265:DNA Helicases; D053484:RecQ Helicases; C106462:Rqh1 protein, S pombe; D004264:DNA Topoisomerases, Type I