Role of Schizosaccharomyces pombe RecQ homolog, recombination, and checkpoint genes in UV damage tolerance (original) (raw)

Abstract

The cellular responses to DNA damage are complex and include direct DNA repair pathways that remove the damage and indirect damage responses which allow cells to survive DNA damage that has not been, or cannot be, removed. We have identified the gene mutated in the rad12.502 strain as a Schizosaccharomyces pombe recQ homolog. The same gene (designated rqh1) is also mutated in the hus2.22 mutant. We show that Rqhl is involved in a DNA damage survival mechanism which prevents cell death when UV-induced DNA damage cannot be removed. This pathway also requires the correct functioning of the recombination machinery and the six checkpoint rad gene products plus the Cdsl kinase. Our data suggest that Rqh1 operates during S phase as part of a mechanism which prevents DNA damage causing cell lethality. This process may involve the bypass of DNA damage sites by the replication fork. Finally, in contrast with the reported literature, we do not find that rqh1 (rad12) mutant cells are defective in UV dimer endonuclease activity.

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Selected References

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  1. Aves S. J., Durkacz B. W., Carr A., Nurse P. Cloning, sequencing and transcriptional control of the Schizosaccharomyces pombe cdc10 'start' gene. EMBO J. 1985 Feb;4(2):457–463. doi: 10.1002/j.1460-2075.1985.tb03651.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barbet N., Muriel W. J., Carr A. M. Versatile shuttle vectors and genomic libraries for use with Schizosaccharomyces pombe. Gene. 1992 May 1;114(1):59–66. doi: 10.1016/0378-1119(92)90707-v. [DOI] [PubMed] [Google Scholar]
  3. Bentley N. J., Holtzman D. A., Flaggs G., Keegan K. S., DeMaggio A., Ford J. C., Hoekstra M., Carr A. M. The Schizosaccharomyces pombe rad3 checkpoint gene. EMBO J. 1996 Dec 2;15(23):6641–6651. [PMC free article] [PubMed] [Google Scholar]
  4. Bowman K. K., Sidik K., Smith C. A., Taylor J. S., Doetsch P. W., Freyer G. A. A new ATP-independent DNA endonuclease from Schizosaccharomyces pombe that recognizes cyclobutane pyrimidine dimers and 6-4 photoproducts. Nucleic Acids Res. 1994 Aug 11;22(15):3026–3032. doi: 10.1093/nar/22.15.3026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carr A. M. Control of cell cycle arrest by the Mec1sc/Rad3sp DNA structure checkpoint pathway. Curr Opin Genet Dev. 1997 Feb;7(1):93–98. doi: 10.1016/s0959-437x(97)80115-3. [DOI] [PubMed] [Google Scholar]
  6. Carr A. M., Hoekstra M. F. The cellular responses to DNA damage. Trends Cell Biol. 1995 Jan;5(1):32–40. doi: 10.1016/s0962-8924(00)88934-5. [DOI] [PubMed] [Google Scholar]
  7. Carr A. M., Sheldrick K. S., Murray J. M., al-Harithy R., Watts F. Z., Lehmann A. R. Evolutionary conservation of excision repair in Schizosaccharomyces pombe: evidence for a family of sequences related to the Saccharomyces cerevisiae RAD2 gene. Nucleic Acids Res. 1993 Mar 25;21(6):1345–1349. doi: 10.1093/nar/21.6.1345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chaganti R. S., Schonberg S., German J. A manyfold increase in sister chromatid exchanges in Bloom's syndrome lymphocytes. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4508–4512. doi: 10.1073/pnas.71.11.4508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chan J. Y., Becker F. F., German J., Ray J. H. Altered DNA ligase I activity in Bloom's syndrome cells. Nature. 1987 Jan 22;325(6102):357–359. doi: 10.1038/325357a0. [DOI] [PubMed] [Google Scholar]
  10. Cimprich K. A., Shin T. B., Keith C. T., Schreiber S. L. cDNA cloning and gene mapping of a candidate human cell cycle checkpoint protein. Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2850–2855. doi: 10.1073/pnas.93.7.2850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ellis N. A., Groden J., Ye T. Z., Straughen J., Lennon D. J., Ciocci S., Proytcheva M., German J. The Bloom's syndrome gene product is homologous to RecQ helicases. Cell. 1995 Nov 17;83(4):655–666. doi: 10.1016/0092-8674(95)90105-1. [DOI] [PubMed] [Google Scholar]
  12. Enoch T., Carr A. M., Nurse P. Fission yeast genes involved in coupling mitosis to completion of DNA replication. Genes Dev. 1992 Nov;6(11):2035–2046. doi: 10.1101/gad.6.11.2035. [DOI] [PubMed] [Google Scholar]
  13. Fernandez Sarabia M. J., McInerny C., Harris P., Gordon C., Fantes P. The cell cycle genes cdc22+ and suc22+ of the fission yeast Schizosaccharomyces pombe encode the large and small subunits of ribonucleotide reductase. Mol Gen Genet. 1993 Apr;238(1-2):241–251. doi: 10.1007/BF00279553. [DOI] [PubMed] [Google Scholar]
  14. Forsburg S. L., Nurse P. The fission yeast cdc19+ gene encodes a member of the MCM family of replication proteins. J Cell Sci. 1994 Oct;107(Pt 10):2779–2788. doi: 10.1242/jcs.107.10.2779. [DOI] [PubMed] [Google Scholar]
  15. Francesconi S., De Recondo A. M., Baldacci G. DNA polymerase delta is required for the replication feedback control of cell cycle progression in Schizosaccharomyces pombe. Mol Gen Genet. 1995 Mar 10;246(5):561–569. doi: 10.1007/BF00298962. [DOI] [PubMed] [Google Scholar]
  16. Freyer G. A., Davey S., Ferrer J. V., Martin A. M., Beach D., Doetsch P. W. An alternative eukaryotic DNA excision repair pathway. Mol Cell Biol. 1995 Aug;15(8):4572–4577. doi: 10.1128/mcb.15.8.4572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gangloff S., McDonald J. P., Bendixen C., Arthur L., Rothstein R. The yeast type I topoisomerase Top3 interacts with Sgs1, a DNA helicase homolog: a potential eukaryotic reverse gyrase. Mol Cell Biol. 1994 Dec;14(12):8391–8398. doi: 10.1128/mcb.14.12.8391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. German J., Bloom D., Passarge E. Bloom's syndrome. VII. Progress report for 1978. Clin Genet. 1979 Apr;15(4):361–367. doi: 10.1111/j.1399-0004.1979.tb01747.x. [DOI] [PubMed] [Google Scholar]
  19. German J., Crippa L. P., Bloom D. Bloom's syndrome. III. Analysis of the chromosome aberration characteristic of this disorder. Chromosoma. 1974;48(4):361–366. doi: 10.1007/BF00290993. [DOI] [PubMed] [Google Scholar]
  20. Gianneli F., Benson P. F., Pawsey S. A., Polani P. E. Ultraviolet light sensitivity and delayed DNA-chain maturation in Bloom's syndrome fibroblasts. Nature. 1977 Feb 3;265(5593):466–469. doi: 10.1038/265466a0. [DOI] [PubMed] [Google Scholar]
  21. Griffiths D. J., Barbet N. C., McCready S., Lehmann A. R., Carr A. M. Fission yeast rad17: a homologue of budding yeast RAD24 that shares regions of sequence similarity with DNA polymerase accessory proteins. EMBO J. 1995 Dec 1;14(23):5812–5823. doi: 10.1002/j.1460-2075.1995.tb00269.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hanada K., Ukita T., Kohno Y., Saito K., Kato J., Ikeda H. RecQ DNA helicase is a suppressor of illegitimate recombination in Escherichia coli. Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3860–3865. doi: 10.1073/pnas.94.8.3860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hand R., German J. A retarded rate of DNA chain growth in Bloom's syndrome. Proc Natl Acad Sci U S A. 1975 Feb;72(2):758–762. doi: 10.1073/pnas.72.2.758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hoheisel J. D., Maier E., Mott R., McCarthy L., Grigoriev A. V., Schalkwyk L. C., Nizetic D., Francis F., Lehrach H. High resolution cosmid and P1 maps spanning the 14 Mb genome of the fission yeast S. pombe. Cell. 1993 Apr 9;73(1):109–120. doi: 10.1016/0092-8674(93)90164-l. [DOI] [PubMed] [Google Scholar]
  25. Kelly T. J., Martin G. S., Forsburg S. L., Stephen R. J., Russo A., Nurse P. The fission yeast cdc18+ gene product couples S phase to START and mitosis. Cell. 1993 Jul 30;74(2):371–382. doi: 10.1016/0092-8674(93)90427-r. [DOI] [PubMed] [Google Scholar]
  26. Langlois R. G., Bigbee W. L., Jensen R. H., German J. Evidence for increased in vivo mutation and somatic recombination in Bloom's syndrome. Proc Natl Acad Sci U S A. 1989 Jan;86(2):670–674. doi: 10.1073/pnas.86.2.670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lehmann A. R., Walicka M., Griffiths D. J., Murray J. M., Watts F. Z., McCready S., Carr A. M. The rad18 gene of Schizosaccharomyces pombe defines a new subgroup of the SMC superfamily involved in DNA repair. Mol Cell Biol. 1995 Dec;15(12):7067–7080. doi: 10.1128/mcb.15.12.7067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lieberman H. B., Hopkins K. M., Nass M., Demetrick D., Davey S. A human homolog of the Schizosaccharomyces pombe rad9+ checkpoint control gene. Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13890–13895. doi: 10.1073/pnas.93.24.13890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lu J., Mullen J. R., Brill S. J., Kleff S., Romeo A. M., Sternglanz R. Human homologues of yeast helicase. Nature. 1996 Oct 24;383(6602):678–679. doi: 10.1038/383678a0. [DOI] [PubMed] [Google Scholar]
  30. MacNeill S. A., Moreno S., Reynolds N., Nurse P., Fantes P. A. The fission yeast Cdc1 protein, a homologue of the small subunit of DNA polymerase delta, binds to Pol3 and Cdc27. EMBO J. 1996 Sep 2;15(17):4613–4628. [PMC free article] [PubMed] [Google Scholar]
  31. Maiorano D., Van Assendelft G. B., Kearsey S. E. Fission yeast cdc21, a member of the MCM protein family, is required for onset of S phase and is located in the nucleus throughout the cell cycle. EMBO J. 1996 Feb 15;15(4):861–872. [PMC free article] [PubMed] [Google Scholar]
  32. Mendonca V. M., Klepin H. D., Matson S. W. DNA helicases in recombination and repair: construction of a delta uvrD delta helD delta recQ mutant deficient in recombination and repair. J Bacteriol. 1995 Mar;177(5):1326–1335. doi: 10.1128/jb.177.5.1326-1335.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Moreno S., Klar A., Nurse P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 1991;194:795–823. doi: 10.1016/0076-6879(91)94059-l. [DOI] [PubMed] [Google Scholar]
  34. Morgan B. A., Conlon F. L., Manzanares M., Millar J. B., Kanuga N., Sharpe J., Krumlauf R., Smith J. C., Sedgwick S. G. Transposon tools for recombinant DNA manipulation: characterization of transcriptional regulators from yeast, Xenopus, and mouse. Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2801–2806. doi: 10.1073/pnas.93.7.2801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Muris D. F., Vreeken K., Carr A. M., Broughton B. C., Lehmann A. R., Lohman P. H., Pastink A. Cloning the RAD51 homologue of Schizosaccharomyces pombe. Nucleic Acids Res. 1993 Sep 25;21(19):4586–4591. doi: 10.1093/nar/21.19.4586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Muris D. F., Vreeken K., Carr A. M., Murray J. M., Smit C., Lohman P. H., Pastink A. Isolation of the Schizosaccharomyces pombe RAD54 homologue, rhp54+, a gene involved in the repair of radiation damage and replication fidelity. J Cell Sci. 1996 Jan;109(Pt 1):73–81. doi: 10.1242/jcs.109.1.73. [DOI] [PubMed] [Google Scholar]
  37. Murray J. M., Doe C. L., Schenk P., Carr A. M., Lehmann A. R., Watts F. Z. Cloning and characterisation of the S. pombe rad15 gene, a homologue to the S. cerevisiae RAD3 and human ERCC2 genes. Nucleic Acids Res. 1992 Jun 11;20(11):2673–2678. doi: 10.1093/nar/20.11.2673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Murray J. M., Tavassoli M., al-Harithy R., Sheldrick K. S., Lehmann A. R., Carr A. M., Watts F. Z. Structural and functional conservation of the human homolog of the Schizosaccharomyces pombe rad2 gene, which is required for chromosome segregation and recovery from DNA damage. Mol Cell Biol. 1994 Jul;14(7):4878–4888. doi: 10.1128/mcb.14.7.4878. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Nasmyth K. A. Temperature-sensitive lethal mutants in the structural gene for DNA ligase in the yeast Schizosaccharomyces pombe. Cell. 1977 Dec;12(4):1109–1120. doi: 10.1016/0092-8674(77)90173-8. [DOI] [PubMed] [Google Scholar]
  40. Nasmyth K., Nurse P. Cell division cycle mutants altered in DNA replication and mitosis in the fission yeast Schizosaccharomyces pombe. Mol Gen Genet. 1981;182(1):119–124. doi: 10.1007/BF00422777. [DOI] [PubMed] [Google Scholar]
  41. Phipps J., Nasim A., Miller D. R. Recovery, repair, and mutagenesis in Schizosaccharomyces pombe. Adv Genet. 1985;23:1–72. doi: 10.1016/s0065-2660(08)60511-8. [DOI] [PubMed] [Google Scholar]
  42. Puranam K. L., Blackshear P. J. Cloning and characterization of RECQL, a potential human homologue of the Escherichia coli DNA helicase RecQ. J Biol Chem. 1994 Nov 25;269(47):29838–29845. [PubMed] [Google Scholar]
  43. Rowley R., Subramani S., Young P. G. Checkpoint controls in Schizosaccharomyces pombe: rad1. EMBO J. 1992 Apr;11(4):1335–1342. doi: 10.1002/j.1460-2075.1992.tb05178.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Russell P., Nurse P. cdc25+ functions as an inducer in the mitotic control of fission yeast. Cell. 1986 Apr 11;45(1):145–153. doi: 10.1016/0092-8674(86)90546-5. [DOI] [PubMed] [Google Scholar]
  45. Savitsky K., Bar-Shira A., Gilad S., Rotman G., Ziv Y., Vanagaite L., Tagle D. A., Smith S., Uziel T., Sfez S. A single ataxia telangiectasia gene with a product similar to PI-3 kinase. Science. 1995 Jun 23;268(5218):1749–1753. doi: 10.1126/science.7792600. [DOI] [PubMed] [Google Scholar]
  46. Seki M., Miyazawa H., Tada S., Yanagisawa J., Yamaoka T., Hoshino S., Ozawa K., Eki T., Nogami M., Okumura K. Molecular cloning of cDNA encoding human DNA helicase Q1 which has homology to Escherichia coli Rec Q helicase and localization of the gene at chromosome 12p12. Nucleic Acids Res. 1994 Nov 11;22(22):4566–4573. doi: 10.1093/nar/22.22.4566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Shinohara A., Ogawa H., Matsuda Y., Ushio N., Ikeo K., Ogawa T. Cloning of human, mouse and fission yeast recombination genes homologous to RAD51 and recA. Nat Genet. 1993 Jul;4(3):239–243. doi: 10.1038/ng0793-239. [DOI] [PubMed] [Google Scholar]
  48. Sidik K., Lieberman H. B., Freyer G. A. Repair of DNA damaged by UV light and ionizing radiation by cell-free extracts prepared from Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):12112–12116. doi: 10.1073/pnas.89.24.12112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Stewart E., Chapman C. R., Al-Khodairy F., Carr A. M., Enoch T. rqh1+, a fission yeast gene related to the Bloom's and Werner's syndrome genes, is required for reversible S phase arrest. EMBO J. 1997 May 15;16(10):2682–2692. doi: 10.1093/emboj/16.10.2682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Takao M., Yonemasu R., Yamamoto K., Yasui A. Characterization of a UV endonuclease gene from the fission yeast Schizosaccharomyces pombe and its bacterial homolog. Nucleic Acids Res. 1996 Apr 1;24(7):1267–1271. doi: 10.1093/nar/24.7.1267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Taylor E. M., McFarlane R. J., Price C. 5-Azacytidine treatment of the fission yeast leads to cytotoxicity and cell cycle arrest. Mol Gen Genet. 1996 Nov 27;253(1-2):128–137. doi: 10.1007/s004380050305. [DOI] [PubMed] [Google Scholar]
  52. Walworth N. C., Bernards R. rad-dependent response of the chk1-encoded protein kinase at the DNA damage checkpoint. Science. 1996 Jan 19;271(5247):353–356. doi: 10.1126/science.271.5247.353. [DOI] [PubMed] [Google Scholar]
  53. Walworth N., Davey S., Beach D. Fission yeast chk1 protein kinase links the rad checkpoint pathway to cdc2. Nature. 1993 May 27;363(6427):368–371. doi: 10.1038/363368a0. [DOI] [PubMed] [Google Scholar]
  54. Watt P. M., Hickson I. D., Borts R. H., Louis E. J. SGS1, a homologue of the Bloom's and Werner's syndrome genes, is required for maintenance of genome stability in Saccharomyces cerevisiae. Genetics. 1996 Nov;144(3):935–945. doi: 10.1093/genetics/144.3.935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Watt P. M., Louis E. J., Borts R. H., Hickson I. D. Sgs1: a eukaryotic homolog of E. coli RecQ that interacts with topoisomerase II in vivo and is required for faithful chromosome segregation. Cell. 1995 Apr 21;81(2):253–260. doi: 10.1016/0092-8674(95)90335-6. [DOI] [PubMed] [Google Scholar]
  56. Willis A. E., Lindahl T. DNA ligase I deficiency in Bloom's syndrome. Nature. 1987 Jan 22;325(6102):355–357. doi: 10.1038/325355a0. [DOI] [PubMed] [Google Scholar]
  57. Yonemasu R., McCready S. J., Murray J. M., Osman F., Takao M., Yamamoto K., Lehmann A. R., Yasui A. Characterization of the alternative excision repair pathway of UV-damaged DNA in Schizosaccharomyces pombe. Nucleic Acids Res. 1997 Apr 15;25(8):1553–1558. doi: 10.1093/nar/25.8.1553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Yu C. E., Oshima J., Fu Y. H., Wijsman E. M., Hisama F., Alisch R., Matthews S., Nakura J., Miki T., Ouais S. Positional cloning of the Werner's syndrome gene. Science. 1996 Apr 12;272(5259):258–262. doi: 10.1126/science.272.5259.258. [DOI] [PubMed] [Google Scholar]
  59. al-Khodairy F., Carr A. M. DNA repair mutants defining G2 checkpoint pathways in Schizosaccharomyces pombe. EMBO J. 1992 Apr;11(4):1343–1350. doi: 10.1002/j.1460-2075.1992.tb05179.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. al-Khodairy F., Fotou E., Sheldrick K. S., Griffiths D. J., Lehmann A. R., Carr A. M. Identification and characterization of new elements involved in checkpoint and feedback controls in fission yeast. Mol Biol Cell. 1994 Feb;5(2):147–160. doi: 10.1091/mbc.5.2.147. [DOI] [PMC free article] [PubMed] [Google Scholar]