Meiotic Recombination Remote from Prominent DNA Break Sites in S. pombe

Young, Jennifer A.; Schreckhise, Randall W.; Steiner, Walter W.; Smith, Gerald R.

doi:10.1016/s1097-2765(02)00452-5

Cited by 116 publications

(227 citation statements)

References 35 publications

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“…In addition, the frequency of meiotic DSB formation in haploid strains is similar to that in diploid strains in pat1-induced meiosis (ref. 17 and this study). Cells were synchronized in G 1 phase of the cell cycle by nitrogen deprivation and shifted to the restrictive temperature of 36°C to induce meiosis.…”

Section: Resultssupporting

confidence: 66%

“…Meiotic DSBs were detected in these cells by Southern blot analysis with a telomere-proximal part of the NotI J fragment of chromosome 1 as a probe ( Fig. 1 A and B) (17). Bands corresponding to the prominent meiotic break site (mbs)1 and mbs2 were detected in pat1 cells in addition to several minor bands ( Fig.…”

Section: Resultsmentioning

confidence: 95%

“…PFGE was performed in 0.8% agarose (chromosomal grade) with a CHEF DR II apparatus (Bio-Rad) for 43-48 h at 2 V͞cm and 8-12°C in TAE buffer (Tris⅐acetate͞EDTA) buffer with a switch time of 30 min. For Southern blot analysis, agarose plugs were prepared, and the DNA was digested with NotI and fractionated by PFGE before hybridization (17). The 32 P-labeled probes (Ϸ1 kb) were prepared by the PCR with Schizosaccharomyces pombe genomic DNA as the template and 20-nt primers.…”

Section: Pulsed-field Gel Electrophoresis (Pfge) and Southern Blot Anmentioning

confidence: 99%

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A checkpoint control linking meiotic S phase and recombination initiation in fission yeast

Tonami

Murakami²,

Shirahige³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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During meiosis, high levels of recombination initiated by DNA double-strand breaks (DSBs) occur only after DNA replication. However, how DSB formation is coupled to DNA replication is unknown. We examined several DNA replication proteins for a role in this coupling in Schizosaccharomyces pombe, and we show that ribonucleotide reductase, the rate-limiting enzyme of deoxyribonucleotide synthesis and the target of the DNA synthesis inhibitor hydroxyurea (HU) is indirectly required for DSB formation linked to DNA replication. However, in cells in which the function of the DNA-replication-checkpoint proteins Rad1p, Rad3p, Rad9p, Rad17p, Rad26p, Hus1p, or Cds1p was compromised, DSB formation occurred at similar frequencies in the absence or presence of HU. The DSBs in the HU-treated mutant cells occurred at normal sites and were associated with recombination. In addition, Cdc2p is apparently not involved in this process. We propose that the sequence of meiotic S phase and initiation of recombination is coordinated by DNA-replication-checkpoint proteins.cell cycle ͉ DNA replication ͉ meiosis ͉ double-strand break formation D uring meiosis, DNA replication occurs, followed by two rounds of chromosome segregation (1, 2). A high level of homologous recombination occurs after meiotic DNA replication but before the first meiotic nuclear division (1, 3-7). Meiotic recombination is required for the proper segregation of homologs at meiosis I, because a chromatid of one homolog recombines with a chromatid of the other homolog, thereby joining homologous chromosomes.It is well established that meiotic recombination in both budding and fission yeasts is initiated by DNA double-strand breaks (DSBs), catalyzed by the DNA topoisomerase type-2-related enzyme Spo11 in budding yeast (8) and Rec12p in fission yeast (6). Both Spo11 in budding yeast and Rec12p in fission yeast are also essential for meiotic recombination. In addition to Rec12p, meiotic DSB requires multiple gene products that are also essential for meiotic recombination in two yeasts (1, 2).The DNA-replication checkpoint operates during both the mitotic cell cycle and meiosis to ensure that chromosome segregation is blocked if DNA replication is incomplete (9-12). The proteins Rad1p, Rad3p, Rad9p, Rad17p, Rad26p, Hus1p, and Cds1p are required for operation of the DNA-replication checkpoint in fission yeast (9-11, 13). The DNA-replicationcheckpoint proteins send a signal to block mitosis via the cyclin-dependent kinase Cdc2p complexed with the B-type cyclin Cdc13p (9). The Cdc2p-Cdc13p protein kinase is fully activated at the onset of mitosis by dephosphorylation of Cdc2p Tyr-15 (9). Phosphorylation of Cdc2p on Tyr-15 is catalyzed by both the Wee1p and Mik1p Tyr kinases, and dephosphorylation is carried out mainly by the Cdc25p Tyr phosphatase (14).It has been proposed that DNA replication is directly coupled to initiation of DSB formation in budding yeast, because inhibition of the completion of DNA replication either by hydroxyurea (HU) treatment or by mutation of both ...

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Section: Resultssupporting

confidence: 66%

Section: Resultsmentioning

confidence: 95%

Section: Pulsed-field Gel Electrophoresis (Pfge) and Southern Blot Anmentioning

confidence: 99%

See 1 more Smart Citation

A checkpoint control linking meiotic S phase and recombination initiation in fission yeast

Tonami

Murakami²,

Shirahige³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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“…To address whether the requirement for Rec10 throughout the genome is specific for crossovers, we also tested gene conversion (cen1, cen2, cen3). The bar is 100 cM based on 0.16 cM/kb (Young et al 2002). frequencies at the lys7 and ura1 loci. lys7 gene conversion levels were greatly reduced in two-factor crosses of rec10-144 and rec10-155 mutants.…”

Section: Resultsmentioning

confidence: 99%

Linear Element-Independent Meiotic Recombination in Schizosaccharomyces pombe

et al. 2006

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Most organisms form protein-rich, linear, ladder-like structures associated with chromosomes during early meiosis, the synaptonemal complex. In Schizosaccharomyces pombe, linear elements (LinEs) are thread-like, proteinacious chromosome-associated structures that form during early meiosis. LinEs are related to axial elements, the synaptonemal complex precursors of other organisms. Previous studies have led to the suggestion that axial structures are essential to mediate meiotic recombination. Rec10 protein is a major component of S. pombe LinEs and is required for their development. In this report we study recombination in a number of rec10 mutants, one of which (rec10-155) does not form LinEs, but is predicted to encode a truncated Rec10 protein. This mutant has levels of crossing over and gene conversion substantially higher than a rec10 null mutant (rec10-175) and forms cytologically detectable Rad51 foci indicative of meiotic recombination intermediates. These data demonstrate that while Rec10 is required for meiotic recombination, substantial meiotic recombination can occur in rec10 mutants that do not form LinEs, indicating that LinEs per se are not essential for all meiotic recombination.

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“…Maybe the K22M activity is sufficient to allow some Mre11 nuclease activity for removing Spo11 from meiotic breaks, whereas the other two S alleles tested were too strongly affected in resection of a diverse spectrum of substrates to be viable. In support of a conservation of the effect of the Fcanonical_ Rad50S mutation is the fact that, when modelled in the fission yeast Schizosaccharomyces pombe, the K81I rad50S mutation was viable and behaved as in S. cerevisiae, showing formation but no resection of meiotic DSB, and thus no viable spores (Young et al 2002). Since Spo11Yoligonucleotide complexes, most probably resulting from endonucleolytic release by the Mre11 complex, have been detected during mouse meiosis (Neale et al 2005), it would be interesting to see if this intermediate exists in the same amounts in the meiotic cells of the different Mre11 complex mutations mouse models, or if there might be an alternative way of processing meiotic DSB in mice.…”

Section: Animal Models For Mutations In the Mre11 Complexmentioning

confidence: 97%

The multiple roles of the Mre11 complex for meiotic recombination

2007

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During the first meiotic prophase, numerous DNA double-strand breaks (DSB) are formed in the genome in order to initiate recombination between homologous chromosomes. The conserved Mre11 complex, formed of Mre11, Rad50 and Nbs1 (Xrs2 in Saccharomyces cerevisiae) proteins, plays a crucial role in mitotic cells for sensing and repairing DSB. In meiosis the Mre11 complex is also required for meiotic recombination. Depending on the organisms, the Mre11 complex is required for the formation of the DSB catalysed by the transesterase Spo11 protein. It then plays a unique function in removing covalently attached Spo11 from the 5 ¶ extremity of the breaks through its nuclease activity, to allow further break resection. Finally, the Mre11 complex also plays a role during meiosis in bridging DNA molecules together and in sensing Spo11 DSB and activating the DNA damage checkpoint. In this article the different biochemical functions of the Mre11 complex required during meiosis are reviewed, as well as the consequences of Mre11 complex inactivation for meiosis in several organisms. Finally, I describe the meiotic phenotypes of several animal models that have been developed to model hypomorphic mutations of the Mre11 complex, involved in humans in some genetic instability disorders.

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Meiotic Recombination Remote from Prominent DNA Break Sites in S. pombe

Cited by 116 publications

References 35 publications

A checkpoint control linking meiotic S phase and recombination initiation in fission yeast

A checkpoint control linking meiotic S phase and recombination initiation in fission yeast

Linear Element-Independent Meiotic Recombination in Schizosaccharomyces pombe

The multiple roles of the Mre11 complex for meiotic recombination

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