Abstract
Most models of homologous recombination invoke cleavage of Holliday junctions to explain crossing over. The Mus81.Eme1 endonuclease from fission yeast and humans cleaves Holliday junctions and other branched DNA structures, leaving its physiological substrate uncertain. We report here that Schizosaccharomyces pombe mus81 mutants have normal or elevated frequencies of gene conversion but 20- to 100-fold reduced frequencies of crossing over. Thus, gene conversion and crossing over can be genetically separated, and Mus81 is required for crossing over, supporting the hypothesis that the fission yeast Mus81.Eme1 protein complex resolves Holliday junctions in meiotic cells.
Full Text
The Full Text of this article is available as a PDF (70.5 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Allers T., Lichten M. Differential timing and control of noncrossover and crossover recombination during meiosis. Cell. 2001 Jul 13;106(1):47–57. doi: 10.1016/s0092-8674(01)00416-0. [DOI] [PubMed] [Google Scholar]
- Benson F. E., West S. C. Substrate specificity of the Escherichia coli RuvC protein. Resolution of three- and four-stranded recombination intermediates. J Biol Chem. 1994 Feb 18;269(7):5195–5201. [PubMed] [Google Scholar]
- Boddy M. N., Gaillard P. H., McDonald W. H., Shanahan P., Yates J. R., 3rd, Russell P. Mus81-Eme1 are essential components of a Holliday junction resolvase. Cell. 2001 Nov 16;107(4):537–548. doi: 10.1016/s0092-8674(01)00536-0. [DOI] [PubMed] [Google Scholar]
- Boddy M. N., Lopez-Girona A., Shanahan P., Interthal H., Heyer W. D., Russell P. Damage tolerance protein Mus81 associates with the FHA1 domain of checkpoint kinase Cds1. Mol Cell Biol. 2000 Dec;20(23):8758–8766. doi: 10.1128/mcb.20.23.8758-8766.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bolt Edward L., Lloyd Robert G. Substrate specificity of RusA resolvase reveals the DNA structures targeted by RuvAB and RecG in vivo. Mol Cell. 2002 Jul;10(1):187–198. doi: 10.1016/s1097-2765(02)00560-9. [DOI] [PubMed] [Google Scholar]
- Bähler J., Wu J. Q., Longtine M. S., Shah N. G., McKenzie A., 3rd, Steever A. B., Wach A., Philippsen P., Pringle J. R. Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Yeast. 1998 Jul;14(10):943–951. doi: 10.1002/(SICI)1097-0061(199807)14:10<943::AID-YEA292>3.0.CO;2-Y. [DOI] [PubMed] [Google Scholar]
- Cervantes M. D., Farah J. A., Smith G. R. Meiotic DNA breaks associated with recombination in S. pombe. Mol Cell. 2000 May;5(5):883–888. doi: 10.1016/s1097-2765(00)80328-7. [DOI] [PubMed] [Google Scholar]
- Chan S. N., Harris L., Bolt E. L., Whitby M. C., Lloyd R. G. Sequence specificity and biochemical characterization of the RusA Holliday junction resolvase of Escherichia coli. J Biol Chem. 1997 Jun 6;272(23):14873–14882. doi: 10.1074/jbc.272.23.14873. [DOI] [PubMed] [Google Scholar]
- Chen X. B., Melchionna R., Denis C. M., Gaillard P. H., Blasina A., Van de Weyer I., Boddy M. N., Russell P., Vialard J., McGowan C. H. Human Mus81-associated endonuclease cleaves Holliday junctions in vitro. Mol Cell. 2001 Nov;8(5):1117–1127. doi: 10.1016/s1097-2765(01)00375-6. [DOI] [PubMed] [Google Scholar]
- Ciccia Alberto, Constantinou Angelos, West Stephen C. Identification and characterization of the human mus81-eme1 endonuclease. J Biol Chem. 2003 Apr 29;278(27):25172–25178. doi: 10.1074/jbc.M302882200. [DOI] [PubMed] [Google Scholar]
- Constantinou A., Davies A. A., West S. C. Branch migration and Holliday junction resolution catalyzed by activities from mammalian cells. Cell. 2001 Jan 26;104(2):259–268. doi: 10.1016/s0092-8674(01)00210-0. [DOI] [PubMed] [Google Scholar]
- Davis L., Smith G. R. Meiotic recombination and chromosome segregation in Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8395–8402. doi: 10.1073/pnas.121005598. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Farah Joseph A., Hartsuiker Edgar, Mizuno Ken-Ichi, Ohta Kunihiro, Smith Gerald R. A 160-bp palindrome is a Rad50.Rad32-dependent mitotic recombination hotspot in Schizosaccharomyces pombe. Genetics. 2002 May;161(1):461–468. doi: 10.1093/genetics/161.1.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gaillard Pierre-Henri L., Noguchi Eishi, Shanahan Paul, Russell Paul. The endogenous Mus81-Eme1 complex resolves Holliday junctions by a nick and counternick mechanism. Mol Cell. 2003 Sep;12(3):747–759. doi: 10.1016/s1097-2765(03)00342-3. [DOI] [PubMed] [Google Scholar]
- 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]
- Interthal H., Heyer W. D. MUS81 encodes a novel helix-hairpin-helix protein involved in the response to UV- and methylation-induced DNA damage in Saccharomyces cerevisiae. Mol Gen Genet. 2000 Jun;263(5):812–827. doi: 10.1007/s004380000241. [DOI] [PubMed] [Google Scholar]
- Kaliraman V., Mullen J. R., Fricke W. M., Bastin-Shanower S. A., Brill S. J. Functional overlap between Sgs1-Top3 and the Mms4-Mus81 endonuclease. Genes Dev. 2001 Oct 15;15(20):2730–2740. doi: 10.1101/gad.932201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lloyd R. G. Conjugational recombination in resolvase-deficient ruvC mutants of Escherichia coli K-12 depends on recG. J Bacteriol. 1991 Sep;173(17):5414–5418. doi: 10.1128/jb.173.17.5414-5418.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McKim K. S., Green-Marroquin B. L., Sekelsky J. J., Chin G., Steinberg C., Khodosh R., Hawley R. S. Meiotic synapsis in the absence of recombination. Science. 1998 Feb 6;279(5352):876–878. doi: 10.1126/science.279.5352.876. [DOI] [PubMed] [Google Scholar]
- McKim K. S., Hayashi-Hagihara A. mei-W68 in Drosophila melanogaster encodes a Spo11 homolog: evidence that the mechanism for initiating meiotic recombination is conserved. Genes Dev. 1998 Sep 15;12(18):2932–2942. doi: 10.1101/gad.12.18.2932. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meselson M. S., Radding C. M. A general model for genetic recombination. Proc Natl Acad Sci U S A. 1975 Jan;72(1):358–361. doi: 10.1073/pnas.72.1.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mizuuchi K., Kemper B., Hays J., Weisberg R. A. T4 endonuclease VII cleaves holliday structures. Cell. 1982 Jun;29(2):357–365. doi: 10.1016/0092-8674(82)90152-0. [DOI] [PubMed] [Google Scholar]
- Mullen J. R., Kaliraman V., Ibrahim S. S., Brill S. J. Requirement for three novel protein complexes in the absence of the Sgs1 DNA helicase in Saccharomyces cerevisiae. Genetics. 2001 Jan;157(1):103–118. doi: 10.1093/genetics/157.1.103. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Munz P. An analysis of interference in the fission yeast Schizosaccharomyces pombe. Genetics. 1994 Jul;137(3):701–707. doi: 10.1093/genetics/137.3.701. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roeder G. S. Meiotic chromosomes: it takes two to tango. Genes Dev. 1997 Oct 15;11(20):2600–2621. doi: 10.1101/gad.11.20.2600. [DOI] [PubMed] [Google Scholar]
- Roman H., Fabre F. Gene conversion and associated reciprocal recombination are separable events in vegetative cells of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6912–6916. doi: 10.1073/pnas.80.22.6912. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Wood V., Gwilliam R., Rajandream M-A, Lyne M., Lyne R., Stewart A., Sgouros J., Peat N., Hayles J., Baker S. The genome sequence of Schizosaccharomyces pombe. Nature. 2002 Feb 21;415(6874):871–880. doi: 10.1038/nature724. [DOI] [PubMed] [Google Scholar]
- Young Jennifer A., Schreckhise Randall W., Steiner Walter W., Smith Gerald R. Meiotic recombination remote from prominent DNA break sites in S. pombe. Mol Cell. 2002 Feb;9(2):253–263. doi: 10.1016/s1097-2765(02)00452-5. [DOI] [PubMed] [Google Scholar]
- de los Santos T., Loidl J., Larkin B., Hollingsworth N. M. A role for MMS4 in the processing of recombination intermediates during meiosis in Saccharomyces cerevisiae. Genetics. 2001 Dec;159(4):1511–1525. doi: 10.1093/genetics/159.4.1511. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de los Santos Teresa, Hunter Neil, Lee Cindy, Larkin Brittany, Loidl Josef, Hollingsworth Nancy M. The Mus81/Mms4 endonuclease acts independently of double-Holliday junction resolution to promote a distinct subset of crossovers during meiosis in budding yeast. Genetics. 2003 May;164(1):81–94. doi: 10.1093/genetics/164.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]