Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1996 Dec;16(12):6783–6793. doi: 10.1128/mcb.16.12.6783

Dual requirement for the yeast MMS19 gene in DNA repair and RNA polymerase II transcription.

S Lauder 1, M Bankmann 1, S N Guzder 1, P Sung 1, L Prakash 1, S Prakash 1
PMCID: PMC231681  PMID: 8943333

Abstract

Genetic and biochemical studies of Saccharomyces cerevisiae have indicated the involvement of a large number of protein factors in nucleotide excision repair (NER) of UV-damaged DNA. However, how MMS19 affects this process has remained unclear. Here, we report on the isolation of the MMS19 gene and the determination of its role in NER and other cellular processes. Genetic and biochemical evidence indicates that besides its function in NER, MMS19 also affects RNA polymerase II (Pol II) transcription. mms19delta cells do not grow at 37 degrees C, and mutant extract exhibits a thermolabile defect in Pol II transcription. Thus, Mms19 protein resembles TFIIH in that it is required for both transcription and DNA repair. However, addition of purified Mms19 protein does not alleviate the transcriptional defect of the mms19delta extract, nor does it stimulate the incision of UV-damaged DNA reconstituted from purified proteins. Interestingly, addition of purified TFIIH corrects the transcriptional defect of the mms19delta extract. Mms19 is, however, not a component of TFIIH or of Pol II holoenzyme. These and other results suggest that Mms19 affects NER and transcription by influencing the activity of TFIIH as an upstream regulatory element. It is proposed that mutations in the human MMS19 counterpart could result in syndromes in which both NER and transcription are affected.

Full Text

The Full Text of this article is available as a PDF (445.1 KB).

Selected References

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

  1. Bailly V., Sommers C. H., Sung P., Prakash L., Prakash S. Specific complex formation between proteins encoded by the yeast DNA repair and recombination genes RAD1 and RAD10. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8273–8277. doi: 10.1073/pnas.89.17.8273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bardwell A. J., Bardwell L., Tomkinson A. E., Friedberg E. C. Specific cleavage of model recombination and repair intermediates by the yeast Rad1-Rad10 DNA endonuclease. Science. 1994 Sep 30;265(5181):2082–2085. doi: 10.1126/science.8091230. [DOI] [PubMed] [Google Scholar]
  3. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Carlson M., Botstein D. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase. Cell. 1982 Jan;28(1):145–154. doi: 10.1016/0092-8674(82)90384-1. [DOI] [PubMed] [Google Scholar]
  6. Culotti J., Hartwell L. H. Genetic control of the cell division cycle in yeast. 3. Seven genes controlling nuclear division. Exp Cell Res. 1971 Aug;67(2):389–401. doi: 10.1016/0014-4827(71)90424-1. [DOI] [PubMed] [Google Scholar]
  7. Feaver W. J., Svejstrup J. Q., Bardwell L., Bardwell A. J., Buratowski S., Gulyas K. D., Donahue T. F., Friedberg E. C., Kornberg R. D. Dual roles of a multiprotein complex from S. cerevisiae in transcription and DNA repair. Cell. 1993 Dec 31;75(7):1379–1387. doi: 10.1016/0092-8674(93)90624-y. [DOI] [PubMed] [Google Scholar]
  8. Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
  9. Guzder S. N., Bailly V., Sung P., Prakash L., Prakash S. Yeast DNA repair protein RAD23 promotes complex formation between transcription factor TFIIH and DNA damage recognition factor RAD14. J Biol Chem. 1995 Apr 14;270(15):8385–8388. doi: 10.1074/jbc.270.15.8385. [DOI] [PubMed] [Google Scholar]
  10. Guzder S. N., Habraken Y., Sung P., Prakash L., Prakash S. Reconstitution of yeast nucleotide excision repair with purified Rad proteins, replication protein A, and transcription factor TFIIH. J Biol Chem. 1995 Jun 2;270(22):12973–12976. doi: 10.1074/jbc.270.22.12973. [DOI] [PubMed] [Google Scholar]
  11. Guzder S. N., Qiu H., Sommers C. H., Sung P., Prakash L., Prakash S. DNA repair gene RAD3 of S. cerevisiae is essential for transcription by RNA polymerase II. Nature. 1994 Jan 6;367(6458):91–94. doi: 10.1038/367091a0. [DOI] [PubMed] [Google Scholar]
  12. Guzder S. N., Sung P., Bailly V., Prakash L., Prakash S. RAD25 is a DNA helicase required for DNA repair and RNA polymerase II transcription. Nature. 1994 Jun 16;369(6481):578–581. doi: 10.1038/369578a0. [DOI] [PubMed] [Google Scholar]
  13. Guzder S. N., Sung P., Prakash L., Prakash S. Nucleotide excision repair in yeast is mediated by sequential assembly of repair factors and not by a pre-assembled repairosome. J Biol Chem. 1996 Apr 12;271(15):8903–8910. doi: 10.1074/jbc.271.15.8903. [DOI] [PubMed] [Google Scholar]
  14. Guzder S. N., Sung P., Prakash L., Prakash S. Yeast DNA-repair gene RAD14 encodes a zinc metalloprotein with affinity for ultraviolet-damaged DNA. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5433–5437. doi: 10.1073/pnas.90.12.5433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Guzder S. N., Sung P., Prakash S., Prakash L. Lethality in yeast of trichothiodystrophy (TTD) mutations in the human xeroderma pigmentosum group D gene. Implications for transcriptional defect in TTD. J Biol Chem. 1995 Jul 28;270(30):17660–17663. doi: 10.1074/jbc.270.30.17660. [DOI] [PubMed] [Google Scholar]
  16. Habraken Y., Sung P., Prakash L., Prakash S. Structure-specific nuclease activity in yeast nucleotide excision repair protein Rad2. J Biol Chem. 1995 Dec 15;270(50):30194–30198. doi: 10.1074/jbc.270.50.30194. [DOI] [PubMed] [Google Scholar]
  17. Habraken Y., Sung P., Prakash L., Prakash S. Yeast excision repair gene RAD2 encodes a single-stranded DNA endonuclease. Nature. 1993 Nov 25;366(6453):365–368. doi: 10.1038/366365a0. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Kingsman S. M., Cousens D., Stanway C. A., Chambers A., Wilson M., Kingsman A. J. High-efficiency yeast expression vectors based on the promoter of the phosphoglycerate kinase gene. Methods Enzymol. 1990;185:329–341. doi: 10.1016/0076-6879(90)85029-n. [DOI] [PubMed] [Google Scholar]
  20. Lopez J. A., Chung D. W., Fujikawa K., Hagen F. S., Papayannopoulou T., Roth G. J. Cloning of the alpha chain of human platelet glycoprotein Ib: a transmembrane protein with homology to leucine-rich alpha 2-glycoprotein. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5615–5619. doi: 10.1073/pnas.84.16.5615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Mu D., Hsu D. S., Sancar A. Reaction mechanism of human DNA repair excision nuclease. J Biol Chem. 1996 Apr 5;271(14):8285–8294. doi: 10.1074/jbc.271.14.8285. [DOI] [PubMed] [Google Scholar]
  23. Mu D., Park C. H., Matsunaga T., Hsu D. S., Reardon J. T., Sancar A. Reconstitution of human DNA repair excision nuclease in a highly defined system. J Biol Chem. 1995 Feb 10;270(6):2415–2418. doi: 10.1074/jbc.270.6.2415. [DOI] [PubMed] [Google Scholar]
  24. Ohkura H., Yanagida M. S. pombe gene sds22+ essential for a midmitotic transition encodes a leucine-rich repeat protein that positively modulates protein phosphatase-1. Cell. 1991 Jan 11;64(1):149–157. doi: 10.1016/0092-8674(91)90216-l. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Prakash L., Prakash S. Three additional genes involved in pyrimidine dimer removal in Saccharomyces cerevisiae: RAD7, RAD14 and MMS19. Mol Gen Genet. 1979 Nov;176(3):351–359. doi: 10.1007/BF00333097. [DOI] [PubMed] [Google Scholar]
  27. Prakash S., Sung P., Prakash L. DNA repair genes and proteins of Saccharomyces cerevisiae. Annu Rev Genet. 1993;27:33–70. doi: 10.1146/annurev.ge.27.120193.000341. [DOI] [PubMed] [Google Scholar]
  28. Qiu H., Park E., Prakash L., Prakash S. The Saccharomyces cerevisiae DNA repair gene RAD25 is required for transcription by RNA polymerase II. Genes Dev. 1993 Nov;7(11):2161–2171. doi: 10.1101/gad.7.11.2161. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sayre M. H., Tschochner H., Kornberg R. D. Reconstitution of transcription with five purified initiation factors and RNA polymerase II from Saccharomyces cerevisiae. J Biol Chem. 1992 Nov 15;267(32):23376–23382. [PubMed] [Google Scholar]
  32. Sung P., Bailly V., Weber C., Thompson L. H., Prakash L., Prakash S. Human xeroderma pigmentosum group D gene encodes a DNA helicase. Nature. 1993 Oct 28;365(6449):852–855. doi: 10.1038/365852a0. [DOI] [PubMed] [Google Scholar]
  33. Sung P., Guzder S. N., Prakash L., Prakash S. Reconstitution of TFIIH and requirement of its DNA helicase subunits, Rad3 and Rad25, in the incision step of nucleotide excision repair. J Biol Chem. 1996 May 3;271(18):10821–10826. doi: 10.1074/jbc.271.18.10821. [DOI] [PubMed] [Google Scholar]
  34. Sung P., Higgins D., Prakash L., Prakash S. Mutation of lysine-48 to arginine in the yeast RAD3 protein abolishes its ATPase and DNA helicase activities but not the ability to bind ATP. EMBO J. 1988 Oct;7(10):3263–3269. doi: 10.1002/j.1460-2075.1988.tb03193.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. 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]
  37. Sweder K. S., Hanawalt P. C. The COOH terminus of suppressor of stem loop (SSL2/RAD25) in yeast is essential for overall genomic excision repair and transcription-coupled repair. J Biol Chem. 1994 Jan 21;269(3):1852–1857. [PubMed] [Google Scholar]
  38. Tomkinson A. E., Bardwell A. J., Bardwell L., Tappe N. J., Friedberg E. C. Yeast DNA repair and recombination proteins Rad1 and Rad10 constitute a single-stranded-DNA endonuclease. Nature. 1993 Apr 29;362(6423):860–862. doi: 10.1038/362860a0. [DOI] [PubMed] [Google Scholar]
  39. Watkins J. F., Sung P., Prakash L., Prakash S. The Saccharomyces cerevisiae DNA repair gene RAD23 encodes a nuclear protein containing a ubiquitin-like domain required for biological function. Mol Cell Biol. 1993 Dec;13(12):7757–7765. doi: 10.1128/mcb.13.12.7757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Woontner M., Wade P. A., Bonner J., Jaehning J. A. Transcriptional activation in an improved whole-cell extract from Saccharomyces cerevisiae. Mol Cell Biol. 1991 Sep;11(9):4555–4560. doi: 10.1128/mcb.11.9.4555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Young D., Riggs M., Field J., Vojtek A., Broek D., Wigler M. The adenylyl cyclase gene from Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7989–7993. doi: 10.1073/pnas.86.20.7989. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES