Sgs1 function in the repair of DNA replication intermediates is separable from its role in homologous recombinational repair.
Mutations in human homologues of the bacterial RecQ helicase cause diseases leading to cancer predisposition and/or shortened lifespan (Werner, Bloom, and Rothmund-Thomson syndromes). The budding yeast Saccharomyces cerevisiae has one RecQ helicase, Sgs1, which functions with Top3 and Rmi1 in DNA repair. Here, we report separation-of-function alleles of SGS1 that suppress the slow growth of top3Delta and rmi1Delta cells similar to an SGS1 deletion, but are resistant to DNA damage similar to wild-type SGS1. In one allele, the second acidic region is deleted, and in the other, only a single aspartic acid residue 664 is deleted. sgs1-D664Delta, unlike sgs1Delta, neither disrupts DNA recombination nor has synthetic growth defects when combined with DNA repair mutants. However, during S phase, it accumulates replication-associated X-shaped structures at damaged replication forks. Furthermore, fluorescent microscopy reveals that the sgs1-D664Delta allele exhibits increased spontaneous RPA foci, suggesting that the persistent X-structures may contain single-stranded DNA. Taken together, these results suggest that the Sgs1 function in repair of DNA replication intermediates can be uncoupled from its role in homologous recombinational repair.
Bernstein
Kara A
KA
Department of Genetics & Development, Columbia University Medical Center, New York, NY 10032, USA.
Shor
Erika
E
Sunjevaric
Ivana
I
Fumasoni
Marco
M
Burgess
Rebecca C
RC
Foiani
Marco
M
Branzei
Dana
D
Rothstein
Rodney
R
D004260:DNA Repair / Q000502:physiology*; D004261:DNA Replication / Q000502:physiology*; D004271:DNA, Fungal / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009154:Mutation; D010641:Phenotype; D053484:RecQ Helicases / Q000235:genetics / Q000502:physiology*; D011995:Recombination, Genetic; D051716:Replication Protein A; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; C082728:RFA1 protein, S cerevisiae; D051716:Replication Protein A; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C090472:SGS1 protein, S cerevisiae; D053484:RecQ Helicases