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. 1988 Sep;8(9):3619–3626. doi: 10.1128/mcb.8.9.3619

RAD1, an excision repair gene of Saccharomyces cerevisiae, is also involved in recombination.

R H Schiestl 1, S Prakash 1
PMCID: PMC365417  PMID: 3065620

Abstract

The RAD1 gene of Saccharomyces cerevisiae is required for the incision step of excision repair of damaged DNA. In this paper, we report our observations on the effect of the RAD1 gene on genetic recombination. Mitotic intrachromosomal and interchromosomal recombination in RAD+, rad1, rad52, and other rad mutant strains was examined. The rad1 deletion mutation and some rad1 point mutations reduced the frequency of intrachromosomal recombination of a his3 duplication, in which one his3 allele is deleted at the 3' end while the other his3 allele is deleted at the 5' end. Mutations in the other excision repair genes, RAD2, RAD3, and RAD4, did not lower recombination frequencies in the his3 duplication. As expected, recombination between the his3 deletion alleles in the duplication was reduced in the rad52 mutant. The frequency of HIS3+ recombinants fell synergistically in the rad1 rad52 double mutant, indicating that the RAD1 and RAD52 genes affect this recombination via different pathways. In contrast to the effect of mutations in the RAD52 gene, mutations in the RAD1 gene did not lower intrachromosomal and interchromosomal recombination between heteroalleles that carry point mutations rather than partial deletions; however, the rad1 delta mutation did lower the frequency of integration of linear plasmids and DNA fragments into homologous genomic sequences. We suggest that RAD1 plays a role in recombination after the formation of the recombinogenic substrate.

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Selected References

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

  1. Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [DOI] [PubMed] [Google Scholar]
  2. Caron P. R., Kushner S. R., Grossman L. Involvement of helicase II (uvrD gene product) and DNA polymerase I in excision mediated by the uvrABC protein complex. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4925–4929. doi: 10.1073/pnas.82.15.4925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dicaprio L., Hastings P. J. Post-meiotic segregation in strains of Saccharomyces cerevisiae unable to excise pyrimidine dimers. Mutat Res. 1976 Oct;37(1):137–140. doi: 10.1016/0027-5107(76)90061-0. [DOI] [PubMed] [Google Scholar]
  4. Dretzen G., Bellard M., Sassone-Corsi P., Chambon P. A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem. 1981 Apr;112(2):295–298. doi: 10.1016/0003-2697(81)90296-7. [DOI] [PubMed] [Google Scholar]
  5. Hartwell L. H., Smith D. Altered fidelity of mitotic chromosome transmission in cell cycle mutants of S. cerevisiae. Genetics. 1985 Jul;110(3):381–395. doi: 10.1093/genetics/110.3.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Higgins D. R., Prakash S., Reynolds P., Polakowska R., Weber S., Prakash L. Isolation and characterization of the RAD3 gene of Saccharomyces cerevisiae and inviability of rad3 deletion mutants. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5680–5684. doi: 10.1073/pnas.80.18.5680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  8. Husain I., Van Houten B., Thomas D. C., Abdel-Monem M., Sancar A. Effect of DNA polymerase I and DNA helicase II on the turnover rate of UvrABC excision nuclease. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6774–6778. doi: 10.1073/pnas.82.20.6774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jackson J. A., Fink G. R. Gene conversion between duplicated genetic elements in yeast. Nature. 1981 Jul 23;292(5821):306–311. doi: 10.1038/292306a0. [DOI] [PubMed] [Google Scholar]
  11. Johnston L. H., Nasmyth K. A. Saccharomyces cerevisiae cell cycle mutant cdc9 is defective in DNA ligase. Nature. 1978 Aug 31;274(5674):891–893. doi: 10.1038/274891a0. [DOI] [PubMed] [Google Scholar]
  12. Kuo C. L., Campbell J. L. Cloning of Saccharomyces cerevisiae DNA replication genes: isolation of the CDC8 gene and two genes that compensate for the cdc8-1 mutation. Mol Cell Biol. 1983 Oct;3(10):1730–1737. doi: 10.1128/mcb.3.10.1730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Madura K., Prakash S. Nucleotide sequence, transcript mapping, and regulation of the RAD2 gene of Saccharomyces cerevisiae. J Bacteriol. 1986 Jun;166(3):914–923. doi: 10.1128/jb.166.3.914-923.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Matson S. W. Escherichia coli helicase II (urvD gene product) translocates unidirectionally in a 3' to 5' direction. J Biol Chem. 1986 Aug 5;261(22):10169–10175. [PubMed] [Google Scholar]
  15. Miller R. D., Prakash L., Prakash S. Defective excision of pyrimidine dimers and interstrand DNA crosslinks in rad7 and rad23 mutants of Saccharomyces cerevisiae. Mol Gen Genet. 1982;188(2):235–239. doi: 10.1007/BF00332681. [DOI] [PubMed] [Google Scholar]
  16. Miller R. D., Prakash L., Prakash S. Genetic control of excision of Saccharomyces cerevisiae interstrand DNA cross-links induced by psoralen plus near-UV light. Mol Cell Biol. 1982 Aug;2(8):939–948. doi: 10.1128/mcb.2.8.939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Montelone B. A., Hoekstra M. F., Malone R. E. Spontaneous mitotic recombination in yeast: the hyper-recombinational rem1 mutations are alleles of the RAD3 gene. Genetics. 1988 Jun;119(2):289–301. doi: 10.1093/genetics/119.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Montelone B. A., Prakash S., Prakash L. Spontaneous mitotic recombination in mms8-1, an allele of the CDC9 gene of Saccharomyces cerevisiae. J Bacteriol. 1981 Aug;147(2):517–525. doi: 10.1128/jb.147.2.517-525.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Naumovski L., Friedberg E. C. A DNA repair gene required for the incision of damaged DNA is essential for viability in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4818–4821. doi: 10.1073/pnas.80.15.4818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Naumovski L., Friedberg E. C. Analysis of the essential and excision repair functions of the RAD3 gene of Saccharomyces cerevisiae by mutagenesis. Mol Cell Biol. 1986 Apr;6(4):1218–1227. doi: 10.1128/mcb.6.4.1218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Naumovski L., Friedberg E. C. The RAD3 gene of Saccharomyces cerevisiae: isolation and characterization of a temperature-sensitive mutant in the essential function and of extragenic suppressors of this mutant. Mol Gen Genet. 1987 Oct;209(3):458–466. doi: 10.1007/BF00331150. [DOI] [PubMed] [Google Scholar]
  22. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354–6358. doi: 10.1073/pnas.78.10.6354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Reynolds P., Higgins D. R., Prakash L., Prakash S. The nucleotide sequence of the RAD3 gene of Saccharomyces cerevisiae: a potential adenine nucleotide binding amino acid sequence and a nonessential acidic carboxyl terminal region. Nucleic Acids Res. 1985 Apr 11;13(7):2357–2372. doi: 10.1093/nar/13.7.2357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Reynolds P., Prakash L., Prakash S. Nucleotide sequence and functional analysis of the RAD1 gene of Saccharomyces cerevisiae. Mol Cell Biol. 1987 Mar;7(3):1012–1020. doi: 10.1128/mcb.7.3.1012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Reynolds R. J., Friedberg E. C. Molecular mechanisms of pyrimidine dimer excision in Saccharomyces cerevisiae: incision of ultraviolet-irradiated deoxyribonucleic acid in vivo. J Bacteriol. 1981 May;146(2):692–704. doi: 10.1128/jb.146.2.692-704.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
  27. Sancar A., Rupp W. D. A novel repair enzyme: UVRABC excision nuclease of Escherichia coli cuts a DNA strand on both sides of the damaged region. Cell. 1983 May;33(1):249–260. doi: 10.1016/0092-8674(83)90354-9. [DOI] [PubMed] [Google Scholar]
  28. Schiestl R. H., Igarashi S., Hastings P. J. Analysis of the mechanism for reversion of a disrupted gene. Genetics. 1988 Jun;119(2):237–247. doi: 10.1093/genetics/119.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schiestl R., Wintersberger U. X-ray enhances mating type switching in heterothallic strains of Saccharomyces cerevisiae. Mol Gen Genet. 1982;186(4):512–517. doi: 10.1007/BF00337958. [DOI] [PubMed] [Google Scholar]
  30. Snow R. Recombination in ultraviolet-sensitive strains of Saccharomyces cerevisiae. Mutat Res. 1968 Nov-Dec;6(3):409–418. doi: 10.1016/0027-5107(68)90058-4. [DOI] [PubMed] [Google Scholar]
  31. Sung P., Prakash L., Matson S. W., Prakash S. RAD3 protein of Saccharomyces cerevisiae is a DNA helicase. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8951–8955. doi: 10.1073/pnas.84.24.8951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sung P., Prakash L., Weber S., Prakash S. The RAD3 gene of Saccharomyces cerevisiae encodes a DNA-dependent ATPase. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6045–6049. doi: 10.1073/pnas.84.17.6045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Szostak J. W., Orr-Weaver T. L., Rothstein R. J., Stahl F. W. The double-strand-break repair model for recombination. Cell. 1983 May;33(1):25–35. doi: 10.1016/0092-8674(83)90331-8. [DOI] [PubMed] [Google Scholar]
  34. Wilcox D. R., Prakash L. Incision and postincision steps of pyrimidine dimer removal in excision-defective mutants of Saccharomyces cerevisiae. J Bacteriol. 1981 Nov;148(2):618–623. doi: 10.1128/jb.148.2.618-623.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yeung A. T., Mattes W. B., Oh E. Y., Grossman L. Enzymatic properties of purified Escherichia coli uvrABC proteins. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6157–6161. doi: 10.1073/pnas.80.20.6157. [DOI] [PMC free article] [PubMed] [Google Scholar]

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