Rad52/Rad59-dependent Recombination as a Means to Rectify Faulty Okazaki Fragment Processing

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The correct removal of 5'-flap structures by Rad27 and Dna2 during Okazaki fragment maturation is crucial for the stable maintenance of genetic materials and cell viability. In this study, we identified RAD52, a key recombination protein, as a multicopy suppressor of dna2-K1080E, a lethal helicase-negative mutant allele of DNA2 in yeasts. In contrast, the overexpression of Rad51, which works conjointly with Rad52 in canonical homologous recombination, failed to suppress the growth defect of the dna2-K1080E mutation, indicating that Rad52 plays a unique and distinct role in Okazaki fragment metabolism. We found that the recombination-defective Rad52QDDD/AAAA mutant did not rescue dna2-K1080E, suggesting that Rad52-mediated recombination is important for suppression. The Rad52-mediated enzymatic stimulation of Dna2 or Rad27 is not a direct cause of suppression observed in vivo, as both Rad52 and Rad52-QDDD/AAAA proteins stimulated the endonuclease activities of both Dna2 and Rad27 to a similar extent. The recombination mediator activity of Rad52 was dispensable for the suppression, whereas both the DNA annealing activity and its ability to interact with Rad59 were essential. In addition, we found that several cohesion establishment factors, including Rsc2 and Elg1, were required for the Rad52-dependent suppression of dna2-K1080E. Our findings suggest a novel Rad52/Rad59-dependent, but Rad51-independent recombination pathway that could ultimately lead to the removal of faulty flaps in conjunction with cohesion establishment factors.
Publisher
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Issue Date
2014-05
Language
English
Article Type
Article
Keywords

REPLICATION PROTEIN-A; DOUBLE-STRAND BREAKS; SISTER-CHROMATID COHESION; YEAST RAD52 PROTEIN; DNA-DAMAGE RESPONSE; ESSENTIAL IN-VIVO; SACCHAROMYCES-CEREVISIAE; HOMOLOGOUS RECOMBINATION; END RESECTION; GENOME STABILITY

Citation

JOURNAL OF BIOLOGICAL CHEMISTRY, v.289, no.21, pp.15064 - 15079

ISSN
0021-9258
DOI
10.1074/jbc.M114.548388
URI
http://hdl.handle.net/10203/190069
Appears in Collection
BS-Journal Papers(저널논문)
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