Skip to main content
PMC home page
Genetics logoLink to Genetics
. 1997 Dec;147(4):1557–1568. doi: 10.1093/genetics/147.4.1557

The Saccharomyces Cerevisiae Rad30 Gene, a Homologue of Escherichia Coli Dinb and Umuc, Is DNA Damage Inducible and Functions in a Novel Error-Free Postreplication Repair Mechanism

J P McDonald 1, A S Levine 1, R Woodgate 1
PMCID: PMC1208331  PMID: 9409821

Abstract

Damage-inducible mutagenesis in prokaryotes is largely dependent upon the activity of the UmuD'C-like proteins. Since many DNA repair processes are structurally and/or functionally conserved between prokaryotes and eukaryotes, we investigated the role of RAD30, a previously uncharacterized Saccharomyces cerevisiae DNA repair gene related to the Escherichia coli dinB, umuC and S. cerevisiae REV1 genes, in UV resistance and UV-induced mutagenesis. Similar to its prokaryotic homologues, RAD30 was found to be damage inducible. Like many S. cerevisiae genes involved in error-prone DNA repair, epistasis analysis clearly places RAD30 in the RAD6 group and rad30 mutants display moderate UV sensitivity reminiscent of rev mutants. However, unlike rev mutants, no defect in UV-induced reversion was seen in rad30 strains. While rad6 and rad18 are both epistatic to rad30, no epistasis was observed with rev1, rev3, rev7 or rad5, all of which are members of the RAD6 epistasis group. These findings suggest that RAD30 participates in a novel error-free repair pathway dependent on RAD6 and RAD18, but independent of REV1, REV3, REV7 and RAD5.

Full Text

The Full Text of this article is available as a PDF (3.7 MB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Bagg A., Kenyon C. J., Walker G. C. Inducibility of a gene product required for UV and chemical mutagenesis in Escherichia coli. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5749–5753. doi: 10.1073/pnas.78.9.5749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bailone A., Sommer S., Knezević J., Dutreix M., Devoret R. A RecA protein mutant deficient in its interaction with the UmuDC complex. Biochimie. 1991 Apr;73(4):479–484. doi: 10.1016/0300-9084(91)90115-h. [DOI] [PubMed] [Google Scholar]
  4. Brotcorne-Lannoye A., Maenhaut-Michel G. Role of RecA protein in untargeted UV mutagenesis of bacteriophage lambda: evidence for the requirement for the dinB gene. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3904–3908. doi: 10.1073/pnas.83.11.3904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bruck I., Woodgate R., McEntee K., Goodman M. F. Purification of a soluble UmuD'C complex from Escherichia coli. Cooperative binding of UmuD'C to single-stranded DNA. J Biol Chem. 1996 May 3;271(18):10767–10774. doi: 10.1074/jbc.271.18.10767. [DOI] [PubMed] [Google Scholar]
  6. Burckhardt S. E., Woodgate R., Scheuermann R. H., Echols H. UmuD mutagenesis protein of Escherichia coli: overproduction, purification, and cleavage by RecA. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1811–1815. doi: 10.1073/pnas.85.6.1811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Echols H., Goodman M. F. Fidelity mechanisms in DNA replication. Annu Rev Biochem. 1991;60:477–511. doi: 10.1146/annurev.bi.60.070191.002401. [DOI] [PubMed] [Google Scholar]
  8. Frank E. G., Hauser J., Levine A. S., Woodgate R. Targeting of the UmuD, UmuD', and MucA' mutagenesis proteins to DNA by RecA protein. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8169–8173. doi: 10.1073/pnas.90.17.8169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Friedberg E. C. Deoxyribonucleic acid repair in the yeast Saccharomyces cerevisiae. Microbiol Rev. 1988 Mar;52(1):70–102. doi: 10.1128/mr.52.1.70-102.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ganesan A. K. Persistence of pyrimidine dimers during post-replication repair in ultraviolet light-irradiated Escherichia coli K12. J Mol Biol. 1974 Jul 25;87(1):103–119. doi: 10.1016/0022-2836(74)90563-4. [DOI] [PubMed] [Google Scholar]
  11. Gietz D., St Jean A., Woods R. A., Schiestl R. H. Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res. 1992 Mar 25;20(6):1425–1425. doi: 10.1093/nar/20.6.1425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Glassner B. J., Mortimer R. K. Synergistic interactions between RAD5, RAD16 and RAD54, three partially homologous yeast DNA repair genes each in a different repair pathway. Radiat Res. 1994 Jul;139(1):24–33. [PubMed] [Google Scholar]
  13. Higgins N. P., Kato K., Strauss B. A model for replication repair in mammalian cells. J Mol Biol. 1976 Mar 5;101(3):417–425. doi: 10.1016/0022-2836(76)90156-x. [DOI] [PubMed] [Google Scholar]
  14. Jang Y. K., Jin Y. H., Shim Y. S., Kim M. J., Yoo E. J., Choi I. S., Lee J. S., Seong R. H., Hong S. H., Park S. D. Identification of the DNA damage-responsive elements of the rhp51+ gene, a recA and RAD51 homolog from the fission yeast Schizosaccharomyces pombe. Mol Gen Genet. 1996 May 23;251(2):167–175. doi: 10.1007/BF02172915. [DOI] [PubMed] [Google Scholar]
  15. Johnson R. E., Henderson S. T., Petes T. D., Prakash S., Bankmann M., Prakash L. Saccharomyces cerevisiae RAD5-encoded DNA repair protein contains DNA helicase and zinc-binding sequence motifs and affects the stability of simple repetitive sequences in the genome. Mol Cell Biol. 1992 Sep;12(9):3807–3818. doi: 10.1128/mcb.12.9.3807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jones J. S., Prakash L. Transcript levels of the Saccharomyces cerevisiae DNA repair gene RAD18 increase in UV irradiated cells and during meiosis but not during the mitotic cell cycle. Nucleic Acids Res. 1991 Feb 25;19(4):893–898. doi: 10.1093/nar/19.4.893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jones J. S., Weber S., Prakash L. The Saccharomyces cerevisiae RAD18 gene encodes a protein that contains potential zinc finger domains for nucleic acid binding and a putative nucleotide binding sequence. Nucleic Acids Res. 1988 Jul 25;16(14B):7119–7131. doi: 10.1093/nar/16.14.7119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Koffel-Schwartz N., Coin F., Veaute X., Fuchs R. P. Cellular strategies for accommodating replication-hindering adducts in DNA: control by the SOS response in Escherichia coli. Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7805–7810. doi: 10.1073/pnas.93.15.7805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kulaeva O. I., Koonin E. V., McDonald J. P., Randall S. K., Rabinovich N., Connaughton J. F., Levine A. S., Woodgate R. Identification of a DinB/UmuC homolog in the archeon Sulfolobus solfataricus. Mutat Res. 1996 Oct 25;357(1-2):245–253. doi: 10.1016/0027-5107(96)00164-9. [DOI] [PubMed] [Google Scholar]
  20. Larimer F. W., Perry J. R., Hardigree A. A. The REV1 gene of Saccharomyces cerevisiae: isolation, sequence, and functional analysis. J Bacteriol. 1989 Jan;171(1):230–237. doi: 10.1128/jb.171.1.230-237.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maenhaut-Michel G., Caillet-Fauquet P. Genetic control of the UV-induced SOS mutator effect in single- and double-stranded DNA phages. Mutat Res. 1990 Jun;230(2):241–254. doi: 10.1016/0027-5107(90)90062-9. [DOI] [PubMed] [Google Scholar]
  22. Morrison A., Christensen R. B., Alley J., Beck A. K., Bernstine E. G., Lemontt J. F., Lawrence C. W. REV3, a Saccharomyces cerevisiae gene whose function is required for induced mutagenesis, is predicted to encode a nonessential DNA polymerase. J Bacteriol. 1989 Oct;171(10):5659–5667. doi: 10.1128/jb.171.10.5659-5667.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nohmi T., Battista J. R., Dodson L. A., Walker G. C. RecA-mediated cleavage activates UmuD for mutagenesis: mechanistic relationship between transcriptional derepression and posttranslational activation. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1816–1820. doi: 10.1073/pnas.85.6.1816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Genetic applications of yeast transformation with linear and gapped plasmids. Methods Enzymol. 1983;101:228–245. doi: 10.1016/0076-6879(83)01017-4. [DOI] [PubMed] [Google Scholar]
  25. Prakash L. Characterization of postreplication repair in Saccharomyces cerevisiae and effects of rad6, rad18, rev3 and rad52 mutations. Mol Gen Genet. 1981;184(3):471–478. doi: 10.1007/BF00352525. [DOI] [PubMed] [Google Scholar]
  26. Resnick M. A., Boyce J., Cox B. Postreplication repair in Saccharomyces cerevisiae. J Bacteriol. 1981 Apr;146(1):285–290. doi: 10.1128/jb.146.1.285-290.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Reynolds P., Weber S., Prakash L. RAD6 gene of Saccharomyces cerevisiae encodes a protein containing a tract of 13 consecutive aspartates. Proc Natl Acad Sci U S A. 1985 Jan;82(1):168–172. doi: 10.1073/pnas.82.1.168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rothstein R. Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. Methods Enzymol. 1991;194:281–301. doi: 10.1016/0076-6879(91)94022-5. [DOI] [PubMed] [Google Scholar]
  29. Rupp W. D., Wilde C. E., 3rd, Reno D. L., Howard-Flanders P. Exchanges between DNA strands in ultraviolet-irradiated Escherichia coli. J Mol Biol. 1971 Oct 14;61(1):25–44. doi: 10.1016/0022-2836(71)90204-x. [DOI] [PubMed] [Google Scholar]
  30. Schmitt M. E., Brown T. A., Trumpower B. L. A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae. Nucleic Acids Res. 1990 May 25;18(10):3091–3092. doi: 10.1093/nar/18.10.3091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shinagawa H., Iwasaki H., Kato T., Nakata A. RecA protein-dependent cleavage of UmuD protein and SOS mutagenesis. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1806–1810. doi: 10.1073/pnas.85.6.1806. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  33. Szekeres E. S., Jr, Woodgate R., Lawrence C. W. Substitution of mucAB or rumAB for umuDC alters the relative frequencies of the two classes of mutations induced by a site-specific T-T cyclobutane dimer and the efficiency of translesion DNA synthesis. J Bacteriol. 1996 May;178(9):2559–2563. doi: 10.1128/jb.178.9.2559-2563.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thomas B. J., Rothstein R. Elevated recombination rates in transcriptionally active DNA. Cell. 1989 Feb 24;56(4):619–630. doi: 10.1016/0092-8674(89)90584-9. [DOI] [PubMed] [Google Scholar]
  35. Torres-Ramos C. A., Yoder B. L., Burgers P. M., Prakash S., Prakash L. Requirement of proliferating cell nuclear antigen in RAD6-dependent postreplicational DNA repair. Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9676–9681. doi: 10.1073/pnas.93.18.9676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wilson R., Ainscough R., Anderson K., Baynes C., Berks M., Bonfield J., Burton J., Connell M., Copsey T., Cooper J. 2.2 Mb of contiguous nucleotide sequence from chromosome III of C. elegans. Nature. 1994 Mar 3;368(6466):32–38. doi: 10.1038/368032a0. [DOI] [PubMed] [Google Scholar]
  37. Woodgate R., Levine A. S. Damage inducible mutagenesis: recent insights into the activities of the Umu family of mutagenesis proteins. Cancer Surv. 1996;28:117–140. [PubMed] [Google Scholar]
  38. Woodgate R., Rajagopalan M., Lu C., Echols H. UmuC mutagenesis protein of Escherichia coli: purification and interaction with UmuD and UmuD'. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7301–7305. doi: 10.1073/pnas.86.19.7301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Xiao W., Singh K. K., Chen B., Samson L. A common element involved in transcriptional regulation of two DNA alkylation repair genes (MAG and MGT1) of Saccharomyces cerevisiae. Mol Cell Biol. 1993 Dec;13(12):7213–7221. doi: 10.1128/mcb.13.12.7213. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES