Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 1997 Aug;146(4):1275–1286. doi: 10.1093/genetics/146.4.1275

Mismatch Repair in Schizosaccharomyces Pombe Requires the Mutl Homologous Gene Pms1: Molecular Cloning and Functional Analysis

P Schar 1, M Baur 1, C Schneider 1, J Kohli 1
PMCID: PMC1208074  PMID: 9258673

Abstract

Homologues of the bacterial mutS and mutL genes involved in DNA mismatch repair have been found in organisms from bacteria to humans. Here, we describe the structure and function of a newly identified Schizosaccharomyces pombe gene that encodes a predicted amino acid sequence of 794 residues with a high degree of homology to MutL related proteins. On the basis of its closer relationship to the eukaryotic ``PMS'' genes than to the ``MLH'' genes, we have designated the S. pombe homologue pms1. Disruption of the pms1 gene causes a significant increase of spontaneous mutagenesis as documented by reversion rate measurements. Tetrad analyses of crosses homozygous for the pms1 mutation reveal a reduction of spore viability from >92% to 80% associated with a low proportion (~50%) of meioses producing four viable spores and a significant, allele-dependent increase of the level of post-meiotic segregation of genetic marker allele pairs. The mutant phenotypes are consistent with a general function of pms1 in correction of mismatched base pairs arising as a consequence of DNA polymerase errors during DNA synthesis, or of hybrid DNA formation between homologous but not perfectly complementary DNA strands during meiotic recombination.

Full Text

The Full Text of this article is available as a PDF (2.3 MB).

Selected References

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

  1. Acharya S., Wilson T., Gradia S., Kane M. F., Guerrette S., Marsischky G. T., Kolodner R., Fishel R. hMSH2 forms specific mispair-binding complexes with hMSH3 and hMSH6. Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13629–13634. doi: 10.1073/pnas.93.24.13629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alani E., Chi N. W., Kolodner R. The Saccharomyces cerevisiae Msh2 protein specifically binds to duplex oligonucleotides containing mismatched DNA base pairs and insertions. Genes Dev. 1995 Jan 15;9(2):234–247. doi: 10.1101/gad.9.2.234. [DOI] [PubMed] [Google Scholar]
  3. Alani E., Reenan R. A., Kolodner R. D. Interaction between mismatch repair and genetic recombination in Saccharomyces cerevisiae. Genetics. 1994 May;137(1):19–39. doi: 10.1093/genetics/137.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Au K. G., Welsh K., Modrich P. Initiation of methyl-directed mismatch repair. J Biol Chem. 1992 Jun 15;267(17):12142–12148. [PubMed] [Google Scholar]
  5. Baker S. M., Bronner C. E., Zhang L., Plug A. W., Robatzek M., Warren G., Elliott E. A., Yu J., Ashley T., Arnheim N. Male mice defective in the DNA mismatch repair gene PMS2 exhibit abnormal chromosome synapsis in meiosis. Cell. 1995 Jul 28;82(2):309–319. doi: 10.1016/0092-8674(95)90318-6. [DOI] [PubMed] [Google Scholar]
  6. Baker S. M., Plug A. W., Prolla T. A., Bronner C. E., Harris A. C., Yao X., Christie D. M., Monell C., Arnheim N., Bradley A. Involvement of mouse Mlh1 in DNA mismatch repair and meiotic crossing over. Nat Genet. 1996 Jul;13(3):336–342. doi: 10.1038/ng0796-336. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Bronner C. E., Baker S. M., Morrison P. T., Warren G., Smith L. G., Lescoe M. K., Kane M., Earabino C., Lipford J., Lindblom A. Mutation in the DNA mismatch repair gene homologue hMLH1 is associated with hereditary non-polyposis colon cancer. Nature. 1994 Mar 17;368(6468):258–261. doi: 10.1038/368258a0. [DOI] [PubMed] [Google Scholar]
  9. Cooper D. L., Lahue R. S., Modrich P. Methyl-directed mismatch repair is bidirectional. J Biol Chem. 1993 Jun 5;268(16):11823–11829. [PubMed] [Google Scholar]
  10. Drummond J. T., Li G. M., Longley M. J., Modrich P. Isolation of an hMSH2-p160 heterodimer that restores DNA mismatch repair to tumor cells. Science. 1995 Jun 30;268(5219):1909–1912. doi: 10.1126/science.7604264. [DOI] [PubMed] [Google Scholar]
  11. Fan J. B., Grothues D., Smith C. L. Alignment of Sfi I sites with the Not I restriction map of Schizosaccharomyces pombe genome. Nucleic Acids Res. 1991 Nov 25;19(22):6289–6294. doi: 10.1093/nar/19.22.6289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fang W. H., Modrich P. Human strand-specific mismatch repair occurs by a bidirectional mechanism similar to that of the bacterial reaction. J Biol Chem. 1993 Jun 5;268(16):11838–11844. [PubMed] [Google Scholar]
  13. Fleck O., Michael H., Heim L. The swi4+ gene of Schizosaccharomyces pombe encodes a homologue of mismatch repair enzymes. Nucleic Acids Res. 1992 May 11;20(9):2271–2278. doi: 10.1093/nar/20.9.2271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fleck O., Schär P., Kohli J. Identification of two mismatch-binding activities in protein extracts of Schizosaccharomyces pombe. Nucleic Acids Res. 1994 Dec 11;22(24):5289–5295. doi: 10.1093/nar/22.24.5289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grimm C., Kohli J., Murray J., Maundrell K. Genetic engineering of Schizosaccharomyces pombe: a system for gene disruption and replacement using the ura4 gene as a selectable marker. Mol Gen Genet. 1988 Dec;215(1):81–86. doi: 10.1007/BF00331307. [DOI] [PubMed] [Google Scholar]
  16. Gutz H. Site Specific Induction of Gene Conversion in SCHIZOSACCHAROMYCES POMBE. Genetics. 1971 Nov;69(3):317–337. doi: 10.1093/genetics/69.3.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Holmes J., Jr, Clark S., Modrich P. Strand-specific mismatch correction in nuclear extracts of human and Drosophila melanogaster cell lines. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5837–5841. doi: 10.1073/pnas.87.15.5837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kohli J., Munz P., Aebi R., Amstutz H., Gysler C., Heyer W. D., Lehmann L., Schuchert P., Szankasi P., Thuriaux P. Interallelic and intergenic conversion in three serine tRNA genes of Schizosaccharomyces pombe. Cold Spring Harb Symp Quant Biol. 1984;49:31–40. doi: 10.1101/sqb.1984.049.01.006. [DOI] [PubMed] [Google Scholar]
  20. Kramer W., Kramer B., Williamson M. S., Fogel S. Cloning and nucleotide sequence of DNA mismatch repair gene PMS1 from Saccharomyces cerevisiae: homology of PMS1 to procaryotic MutL and HexB. J Bacteriol. 1989 Oct;171(10):5339–5346. doi: 10.1128/jb.171.10.5339-5346.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Leach F. S., Nicolaides N. C., Papadopoulos N., Liu B., Jen J., Parsons R., Peltomäki P., Sistonen P., Aaltonen L. A., Nyström-Lahti M. Mutations of a mutS homolog in hereditary nonpolyposis colorectal cancer. Cell. 1993 Dec 17;75(6):1215–1225. doi: 10.1016/0092-8674(93)90330-s. [DOI] [PubMed] [Google Scholar]
  22. Li G. M., Modrich P. Restoration of mismatch repair to nuclear extracts of H6 colorectal tumor cells by a heterodimer of human MutL homologs. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):1950–1954. doi: 10.1073/pnas.92.6.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Maier E., Hoheisel J. D., McCarthy L., Mott R., Grigoriev A. V., Monaco A. P., Larin Z., Lehrach H. Complete coverage of the Schizosaccharomyces pombe genome in yeast artificial chromosomes. Nat Genet. 1992 Jul;1(4):273–277. doi: 10.1038/ng0792-273. [DOI] [PubMed] [Google Scholar]
  24. Marsischky G. T., Filosi N., Kane M. F., Kolodner R. Redundancy of Saccharomyces cerevisiae MSH3 and MSH6 in MSH2-dependent mismatch repair. Genes Dev. 1996 Feb 15;10(4):407–420. doi: 10.1101/gad.10.4.407. [DOI] [PubMed] [Google Scholar]
  25. Modrich P. Mechanisms and biological effects of mismatch repair. Annu Rev Genet. 1991;25:229–253. doi: 10.1146/annurev.ge.25.120191.001305. [DOI] [PubMed] [Google Scholar]
  26. Munz P. On some properties of five mutator alleles in Schizosaccharomyces pombe. Mutat Res. 1975 Jul;29(1):155–157. doi: 10.1016/0027-5107(75)90030-5. [DOI] [PubMed] [Google Scholar]
  27. New L., Liu K., Crouse G. F. The yeast gene MSH3 defines a new class of eukaryotic MutS homologues. Mol Gen Genet. 1993 May;239(1-2):97–108. doi: 10.1007/BF00281607. [DOI] [PubMed] [Google Scholar]
  28. Nicolaides N. C., Papadopoulos N., Liu B., Wei Y. F., Carter K. C., Ruben S. M., Rosen C. A., Haseltine W. A., Fleischmann R. D., Fraser C. M. Mutations of two PMS homologues in hereditary nonpolyposis colon cancer. Nature. 1994 Sep 1;371(6492):75–80. doi: 10.1038/371075a0. [DOI] [PubMed] [Google Scholar]
  29. Nicolas A., Petes T. D. Polarity of meiotic gene conversion in fungi: contrasting views. Experientia. 1994 Mar 15;50(3):242–252. doi: 10.1007/BF01924007. [DOI] [PubMed] [Google Scholar]
  30. Palombo F., Gallinari P., Iaccarino I., Lettieri T., Hughes M., D'Arrigo A., Truong O., Hsuan J. J., Jiricny J. GTBP, a 160-kilodalton protein essential for mismatch-binding activity in human cells. Science. 1995 Jun 30;268(5219):1912–1914. doi: 10.1126/science.7604265. [DOI] [PubMed] [Google Scholar]
  31. Papadopoulos N., Nicolaides N. C., Wei Y. F., Ruben S. M., Carter K. C., Rosen C. A., Haseltine W. A., Fleischmann R. D., Fraser C. M., Adams M. D. Mutation of a mutL homolog in hereditary colon cancer. Science. 1994 Mar 18;263(5153):1625–1629. doi: 10.1126/science.8128251. [DOI] [PubMed] [Google Scholar]
  32. Ponticelli A. S., Sena E. P., Smith G. R. Genetic and physical analysis of the M26 recombination hotspot of Schizosaccharomyces pombe. Genetics. 1988 Jul;119(3):491–497. doi: 10.1093/genetics/119.3.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Prabhala G., Rosenberg G. H., Käufer N. F. Architectural features of pre-mRNA introns in the fission yeast Schizosaccharomyces pombe. Yeast. 1992 Mar;8(3):171–182. doi: 10.1002/yea.320080303. [DOI] [PubMed] [Google Scholar]
  34. Prolla T. A., Christie D. M., Liskay R. M. Dual requirement in yeast DNA mismatch repair for MLH1 and PMS1, two homologs of the bacterial mutL gene. Mol Cell Biol. 1994 Jan;14(1):407–415. doi: 10.1128/mcb.14.1.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Prolla T. A., Pang Q., Alani E., Kolodner R. D., Liskay R. M. MLH1, PMS1, and MSH2 interactions during the initiation of DNA mismatch repair in yeast. Science. 1994 Aug 19;265(5175):1091–1093. doi: 10.1126/science.8066446. [DOI] [PubMed] [Google Scholar]
  36. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  37. Prudhomme M., Martin B., Mejean V., Claverys J. P. Nucleotide sequence of the Streptococcus pneumoniae hexB mismatch repair gene: homology of HexB to MutL of Salmonella typhimurium and to PMS1 of Saccharomyces cerevisiae. J Bacteriol. 1989 Oct;171(10):5332–5338. doi: 10.1128/jb.171.10.5332-5338.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Radman M., Wagner R. Mismatch recognition in chromosomal interactions and speciation. Chromosoma. 1993 Jun;102(6):369–373. doi: 10.1007/BF00360400. [DOI] [PubMed] [Google Scholar]
  39. Reenan R. A., Kolodner R. D. Characterization of insertion mutations in the Saccharomyces cerevisiae MSH1 and MSH2 genes: evidence for separate mitochondrial and nuclear functions. Genetics. 1992 Dec;132(4):975–985. doi: 10.1093/genetics/132.4.975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Reenan R. A., Kolodner R. D. Isolation and characterization of two Saccharomyces cerevisiae genes encoding homologs of the bacterial HexA and MutS mismatch repair proteins. Genetics. 1992 Dec;132(4):963–973. doi: 10.1093/genetics/132.4.963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schär P., Kohli J. Preferential strand transfer and hybrid DNA formation at the recombination hotspot ade6-M26 of Schizosaccharomyces pombe. EMBO J. 1994 Nov 1;13(21):5212–5219. doi: 10.1002/j.1460-2075.1994.tb06852.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schär P., Munz P., Kohli J. Meiotic mismatch repair quantified on the basis of segregation patterns in Schizosaccharomyces pombe. Genetics. 1993 Apr;133(4):815–824. doi: 10.1093/genetics/133.4.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Smith G. R. Hotspots of homologous recombination. Experientia. 1994 Mar 15;50(3):234–241. doi: 10.1007/BF01924006. [DOI] [PubMed] [Google Scholar]
  44. Snow R. Maximum likelihood estimation of linkage and interference from tetrad data. Genetics. 1979 May;92(1):231–245. doi: 10.1093/genetics/92.1.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Strand M., Prolla T. A., Liskay R. M., Petes T. D. Destabilization of tracts of simple repetitive DNA in yeast by mutations affecting DNA mismatch repair. Nature. 1993 Sep 16;365(6443):274–276. doi: 10.1038/365274a0. [DOI] [PubMed] [Google Scholar]
  46. Szankasi P., Heyer W. D., Schuchert P., Kohli J. DNA sequence analysis of the ade6 gene of Schizosaccharomyces pombe. Wild-type and mutant alleles including the recombination host spot allele ade6-M26. J Mol Biol. 1988 Dec 20;204(4):917–925. doi: 10.1016/0022-2836(88)90051-4. [DOI] [PubMed] [Google Scholar]
  47. Szankasi P., Smith G. R. A role for exonuclease I from S. pombe in mutation avoidance and mismatch correction. Science. 1995 Feb 24;267(5201):1166–1169. doi: 10.1126/science.7855597. [DOI] [PubMed] [Google Scholar]
  48. 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]
  49. Tsui H. T., Mandavilli B. S., Winkler M. E. Nonconserved segment of the MutL protein from Escherichia coli K-12 and Salmonella typhimurium. Nucleic Acids Res. 1992 May 11;20(9):2379–2379. doi: 10.1093/nar/20.9.2379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Williamson M. S., Game J. C., Fogel S. Meiotic gene conversion mutants in Saccharomyces cerevisiae. I. Isolation and characterization of pms1-1 and pms1-2. Genetics. 1985 Aug;110(4):609–646. doi: 10.1093/genetics/110.4.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wright A. P., Maundrell K., Shall S. Transformation of Schizosaccharomyces pombe by non-homologous, unstable integration of plasmids in the genome. Curr Genet. 1986;10(7):503–508. doi: 10.1007/BF00447383. [DOI] [PubMed] [Google Scholar]
  52. de Massy B., Rocco V., Nicolas A. The nucleotide mapping of DNA double-strand breaks at the CYS3 initiation site of meiotic recombination in Saccharomyces cerevisiae. EMBO J. 1995 Sep 15;14(18):4589–4598. doi: 10.1002/j.1460-2075.1995.tb00138.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. de Wind N., Dekker M., Berns A., Radman M., te Riele H. Inactivation of the mouse Msh2 gene results in mismatch repair deficiency, methylation tolerance, hyperrecombination, and predisposition to cancer. Cell. 1995 Jul 28;82(2):321–330. doi: 10.1016/0092-8674(95)90319-4. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES