Skip to main content
PMC home page
Genetics logoLink to Genetics
. 1999 Oct;153(2):681–692. doi: 10.1093/genetics/153.2.681

A core activity associated with the N terminus of the yeast RAD52 protein is revealed by RAD51 overexpression suppression of C-terminal rad52 truncation alleles.

E N Asleson 1, R J Okagaki 1, D M Livingston 1
PMCID: PMC1460769  PMID: 10511548

Abstract

C-terminal rad52 truncation and internal deletion mutants were characterized for their ability to repair MMS-induced double-strand breaks and to produce viable spores during meiosis. The rad52-Delta251 allele, encoding the N-terminal 251 amino acids of the predicted 504-amino-acid polypeptide, supports partial activity for both functions. Furthermore, RAD51 overexpression completely suppresses the MMS sensitivity of a rad52-Delta251 mutant. The absence of the C terminus in the truncated protein makes it likely that suppression occurs by bypassing the C-terminal functions of Rad52p. RAD51 overexpression does not suppress the low level of spore viability that the rad52-Delta251 allele causes and only partially suppresses the defect in rad52 alleles encoding the N-terminal 292 or 327 amino acids. The results of this study also show that intragenic complementation between rad52 alleles is governed by a complex relationship that depends heavily on the two alleles involved and their relative dosage. In heteroallelic rad52 diploids, the rad52-Delta251 allele does not complement rad52 missense mutations altering residues 61 or 64 in the N terminus. However, complementation is achieved with each of these missense alleles when the rad52-Delta251 allele is overexpressed. Complementation also occurs between rad52-Delta327 and an internal deletion allele missing residues 210 through 327. We suggest that the first 251 amino acids of Rad52p constitute a core domain that provides critical RAD52 activities.

Full Text

The Full Text of this article is available as a PDF (171.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bai Y., Symington L. S. A Rad52 homolog is required for RAD51-independent mitotic recombination in Saccharomyces cerevisiae. Genes Dev. 1996 Aug 15;10(16):2025–2037. doi: 10.1101/gad.10.16.2025. [DOI] [PubMed] [Google Scholar]
  2. Benson F. E., Baumann P., West S. C. Synergistic actions of Rad51 and Rad52 in recombination and DNA repair. Nature. 1998 Jan 22;391(6665):401–404. doi: 10.1038/34937. [DOI] [PubMed] [Google Scholar]
  3. Boundy-Mills K. L., Livingston D. M. A Saccharomyces cerevisiae RAD52 allele expressing a C-terminal truncation protein: activities and intragenic complementation of missense mutations. Genetics. 1993 Jan;133(1):39–49. doi: 10.1093/genetics/133.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Christianson T. W., Sikorski R. S., Dante M., Shero J. H., Hieter P. Multifunctional yeast high-copy-number shuttle vectors. Gene. 1992 Jan 2;110(1):119–122. doi: 10.1016/0378-1119(92)90454-w. [DOI] [PubMed] [Google Scholar]
  5. Dornfeld K. J., Livingston D. M. Effects of controlled RAD52 expression on repair and recombination in Saccharomyces cerevisiae. Mol Cell Biol. 1991 Apr;11(4):2013–2017. doi: 10.1128/mcb.11.4.2013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Firmenich A. A., Elias-Arnanz M., Berg P. A novel allele of Saccharomyces cerevisiae RFA1 that is deficient in recombination and repair and suppressible by RAD52. Mol Cell Biol. 1995 Mar;15(3):1620–1631. doi: 10.1128/mcb.15.3.1620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hays S. L., Firmenich A. A., Massey P., Banerjee R., Berg P. Studies of the interaction between Rad52 protein and the yeast single-stranded DNA binding protein RPA. Mol Cell Biol. 1998 Jul;18(7):4400–4406. doi: 10.1128/mcb.18.7.4400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Katz W. S., Solomon F. Diversity among beta-tubulins: a carboxy-terminal domain of yeast beta-tubulin is not essential in vivo. Mol Cell Biol. 1988 Jul;8(7):2730–2736. doi: 10.1128/mcb.8.7.2730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kaytor M. D., Livingston D. M. Allele-specific suppression of temperature-sensitive mutations of the Saccharomyces cerevisiae RAD52 gene. Curr Genet. 1996 Feb;29(3):203–210. doi: 10.1007/BF02221549. [DOI] [PubMed] [Google Scholar]
  10. Kaytor M. D., Livingston D. M. Saccharomyces cerevisiae RAD52 alleles temperature-sensitive for the repair of DNA double-strand breaks. Genetics. 1994 Aug;137(4):933–944. doi: 10.1093/genetics/137.4.933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Milne G. T., Weaver D. T. Dominant negative alleles of RAD52 reveal a DNA repair/recombination complex including Rad51 and Rad52. Genes Dev. 1993 Sep;7(9):1755–1765. doi: 10.1101/gad.7.9.1755. [DOI] [PubMed] [Google Scholar]
  12. Mortensen U. H., Bendixen C., Sunjevaric I., Rothstein R. DNA strand annealing is promoted by the yeast Rad52 protein. Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10729–10734. doi: 10.1073/pnas.93.20.10729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. New J. H., Sugiyama T., Zaitseva E., Kowalczykowski S. C. Rad52 protein stimulates DNA strand exchange by Rad51 and replication protein A. Nature. 1998 Jan 22;391(6665):407–410. doi: 10.1038/34950. [DOI] [PubMed] [Google Scholar]
  14. Nguyen M. M., Livingston D. M. Cold-sensitive rad52 alleles of yeast. Curr Genet. 1997 Aug;32(2):100–107. doi: 10.1007/s002940050253. [DOI] [PubMed] [Google Scholar]
  15. Passy S. I., Yu X., Li Z., Radding C. M., Egelman E. H. Rings and filaments of beta protein from bacteriophage lambda suggest a superfamily of recombination proteins. Proc Natl Acad Sci U S A. 1999 Apr 13;96(8):4279–4284. doi: 10.1073/pnas.96.8.4279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Prakash L., Prakash S. Isolation and characterization of MMS-sensitive mutants of Saccharomyces cerevisiae. Genetics. 1977 May;86(1):33–55. doi: 10.1093/genetics/86.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Scherer S., Davis R. W. Replacement of chromosome segments with altered DNA sequences constructed in vitro. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4951–4955. doi: 10.1073/pnas.76.10.4951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schild D. Suppression of a new allele of the yeast RAD52 gene by overexpression of RAD51, mutations in srs2 and ccr4, or mating-type heterozygosity. Genetics. 1995 May;140(1):115–127. doi: 10.1093/genetics/140.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shen Z., Peterson S. R., Comeaux J. C., Zastrow D., Moyzis R. K., Bradbury E. M., Chen D. J. Self-association of human RAD52 protein. Mutat Res. 1996 Oct 18;364(2):81–89. doi: 10.1016/0921-8777(96)00025-0. [DOI] [PubMed] [Google Scholar]
  20. Shinohara A., Ogawa T. Stimulation by Rad52 of yeast Rad51-mediated recombination. Nature. 1998 Jan 22;391(6665):404–407. doi: 10.1038/34943. [DOI] [PubMed] [Google Scholar]
  21. Shinohara A., Shinohara M., Ohta T., Matsuda S., Ogawa T. Rad52 forms ring structures and co-operates with RPA in single-strand DNA annealing. Genes Cells. 1998 Mar;3(3):145–156. doi: 10.1046/j.1365-2443.1998.00176.x. [DOI] [PubMed] [Google Scholar]
  22. Sung P. Function of yeast Rad52 protein as a mediator between replication protein A and the Rad51 recombinase. J Biol Chem. 1997 Nov 7;272(45):28194–28197. doi: 10.1074/jbc.272.45.28194. [DOI] [PubMed] [Google Scholar]
  23. Tyers M., Tokiwa G., Futcher B. Comparison of the Saccharomyces cerevisiae G1 cyclins: Cln3 may be an upstream activator of Cln1, Cln2 and other cyclins. EMBO J. 1993 May;12(5):1955–1968. doi: 10.1002/j.1460-2075.1993.tb05845.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tyers M., Tokiwa G., Nash R., Futcher B. The Cln3-Cdc28 kinase complex of S. cerevisiae is regulated by proteolysis and phosphorylation. EMBO J. 1992 May;11(5):1773–1784. doi: 10.1002/j.1460-2075.1992.tb05229.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Van Dyck E., Hajibagheri N. M., Stasiak A., West S. C. Visualisation of human rad52 protein and its complexes with hRad51 and DNA. J Mol Biol. 1998 Dec 11;284(4):1027–1038. doi: 10.1006/jmbi.1998.2203. [DOI] [PubMed] [Google Scholar]
  26. Van Dyck E., Stasiak A. Z., Stasiak A., West S. C. Binding of double-strand breaks in DNA by human Rad52 protein. Nature. 1999 Apr 22;398(6729):728–731. doi: 10.1038/19560. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES