Identification of joint molecules that form frequently between homologs but rarely between sister chromatids during yeast meiosis

Schwacha, Anthony; Kleckner, Nancy

doi:10.1016/0092-8674(94)90172-4

Cited by 349 publications

(316 citation statements)

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“…Isolated genomic DNA was precipitated in ethanol and dried at 4°C. DNA pellets were dissolved in 50 mM Tris-HCl and 1 mM EDTA (Schwacha and Kleckner, 1994). DNA concentration was measured using the Picogreen assay kit (Invitrogen).…”

Section: Genomic Dna Extractionmentioning

confidence: 99%

“…DNA samples were loaded onto 0.6% Seakem LE agarose gel in TBE buffer at ~2 V/cM for 24 h. Two-dimensional (2D) gel electrophoresis was performed as described elsewhere (Kim et al, 2010;Schwacha and Kleckner, 1994). Genomic DNA was digested as described above, and loaded onto 0.4% Seakem Gold agarose gel without ethidium bromide in TBE buffer at ~1 V/cM for 21 h. Gels were stained with 0.5 μg/ml ethidium bromide for 30 min.…”

Section: Dna Physical Analysismentioning

confidence: 99%

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Shu1 Promotes Homolog Bias of Meiotic Recombination in Saccharomyces cerevisiae

Hong

Kim

2013

Molecules and Cells

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Homologous recombination occurs closely between homologous chromatids with highly ordered recombinosomes through RecA homologs and mediators. The present study demonstrates this relationship during the period of "partner choice" in yeast meiotic recombination. We have examined the formation of recombination intermediates in the absence or presence of Shu1, a member of the PCSS complex, which also includes Psy3, Csm2, and Shu2. DNA physical analysis indicates that Shu1 is essential for promoting the establishment of homolog bias during meiotic homologous recombination, and the partner choice is switched by Mek1 kinase activity. Furthermore, Shu1 promotes both crossover (CO) and non-crossover (NCO) pathways of meiotic recombination. The inactivation of Mek1 kinase allows for meiotic recombination to progress efficiently, but is lost in homolog bias where most doublestrand breaks (DSBs) are repaired via stable intersister joint molecules. Moreover, the Srs2 helicase deletion cells in the budding yeast show slightly reduced COs and NCOs, and Shu1 promotes homolog bias independent of Srs2. Our findings reveal that Shu1 and Mek1 kinase activity have biochemically distinct roles in partner choice, which in turn enhances the understanding of the mechanism associated with the precondition for homolog bias.

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Section: Genomic Dna Extractionmentioning

confidence: 99%

Section: Dna Physical Analysismentioning

confidence: 99%

Shu1 Promotes Homolog Bias of Meiotic Recombination in Saccharomyces cerevisiae

Hong

Kim

2013

Molecules and Cells

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“…Finally, in vivo experiments suggest another accessory protein, Hop2-Mnd1, stimulates Dmc1-dependent and not Rad51-dependent recombination in vivo (Tsubouchi and Roeder 2003;Chen et al 2004;Henry et al 2006). Although these accessory factor differences may allow for distinct regulation of the two recombinases, we reiterate that the ability of each of the recombinases to promote substantial strand invasion in the absence of the other indicates a significant degree of functional redundancy in vivo (Schwacha and Kleckner 1997;Shinohara et al 1997;Zenvirth et al 1997;Bishop et al 1999;Tsubouchi and Roeder 2003).…”

mentioning

confidence: 98%

“…The assembly of the synaptonemal complex brings an axial element with its pair of sisters into close parallel alignment with another axial element holding the homologous sister pair. Three interacting axis-associated proteins-Red1, Hop1, and Mek1-regulate recombination by suppressing Dmc1-independent recombination and, in the case of Red1 and Hop1, by enhancing the efficiency of DSB formation in a locus-specific manner (Rockmill and Roeder 1990;Mao-Draayer et al 1996;Schwacha and Kleckner 1997;Pecina et al 2002). Red, Hop1, and Mek1 also influence the choice of recombination partner (as discussed in more detail below).…”

mentioning

confidence: 99%

“…Previous work showed that Dmc1-independent recombination is inhibited by proteins associated with axial elements (Schwacha and Kleckner 1997;Xu et al 1997;Bishop et al 1999;Niu et al 2005) proteinaceous structures that organize pairs of sister chromatids into two parallel sets of loops by binding at their base. The assembly of the synaptonemal complex brings an axial element with its pair of sisters into close parallel alignment with another axial element holding the homologous sister pair.…”

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confidence: 99%

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Red-Hed regulation: recombinase Rad51, though capable of playing the leading role, may be relegated to supporting Dmc1 in budding yeast meiosis: Figure 1.

Sheridan

Bishop²

2006

Genes Dev.

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DNA strand exchange is the central process of homologous recombination. In budding yeast, this reaction is catalyzed by the recombinases Rad51 and Dmc1. Rad51 is responsible for recombinational repair of DNA damage during mitosis and is also important during meiotic recombination. Dmc1 is only expressed during meiosis and is required for meiotic recombination. The discovery that these two different recombinases are needed for normal levels of meiotic recombination in budding yeast raised the question of how the functions of these proteins relate to one another. The paper by Tsubouchi and Roeder (2006) in this issue of Genes & Development represents important progress toward what is apparently a rather complex answer. Tsubouchi and Roeder (2006) report discovery of a novel meiosis-specific protein, Hed1, which appears to inhibit Rad51 when Dmc1 is absent. The authors show mutation of the HED1 gene allows cells to bypass the meiotic arrest caused by a dmc1 mutation and resolve meiosis-specific double-strand breaks (DSBs) using Rad51. A hed1 mutation can also suppress the defects caused by mutation of HOP2, which codes for a Dmc1 accessory factor. Two-hybrid experiments show that Hed1 and Rad51 can interact directly and immuno-staining shows that subnuclear foci formed by Hed1 and Rad51 colocalize. Together these results imply that Hed1's influence on Rad51 activity is direct. Furthermore, expression of Hed1 in mitotic cells provided evidence that Hed1 expression can be sufficient to inhibit Rad51-dependent repair of mitotic DNA damage.A reasonable interpretation of these results is that the meiotic recombination machinery of budding yeast is regulated such that Dmc1 carries out most strand exchange. Tsubouchi and Roeder's (2006) results also add to evidence indicating that Rad51 is capable of replacing Dmc1's function under certain circumstances. Here, we place these new findings in the context of previous observations and present a model to explain how Rad51 and Dmc1 may cooperate to promote interhomolog recombination in budding yeast, while at the same time accounting for the observed functional redundancy. We further speculate as to how the functional relationship of Rad51 and Dmc1 may have evolved.The notion that Rad51 activity is blocked during meiosis is not new. Previous work showed that Dmc1-independent recombination is inhibited by proteins associated with axial elements (Schwacha and Kleckner 1997;Xu et al. 1997;Bishop et al. 1999;Niu et al. 2005) proteinaceous structures that organize pairs of sister chromatids into two parallel sets of loops by binding at their base. The assembly of the synaptonemal complex brings an axial element with its pair of sisters into close parallel alignment with another axial element holding the homologous sister pair. Three interacting axis-associated proteins-Red1, Hop1, and Mek1-regulate recombination by suppressing Dmc1-independent recombination and, in the case of Red1 and Hop1, by enhancing the efficiency of DSB formation in a locus-specific manner (Rockmill and Roeder...

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Meiotic versus mitotic recombination: Two different routes for double‐strand break repair

2010

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Summary Studies in the yeast Saccharomyces cerevisiae have validated the major features of the double-strand break repair (DSBR) model as an accurate representation of the pathway through which meiotic crossovers are produced. This success has led to this model being invoked to explain double-strand break (DSB) repair in other contexts. However, most non-crossover recombinants generated during S. cerevisiae meiosis do not arise via a DSBR pathway. Furthermore, and it is becoming increasing clear that DSBR is a minor pathway for recombinational repair of DSBs that occur in mitotically proliferating cells; rather, the synthesis-dependent strand annealing (SDSA) model appears to describe mitotic DSB repair more accurately. Fundamental dissimilarities between meiotic and mitotic recombination are not unexpected, since meiotic recombination serves a very different purpose (accurate chromosome segregation, which requires crossovers) than mitotic recombination (repair of DNA damage, which typically generates non-crossovers).

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Identification of joint molecules that form frequently between homologs but rarely between sister chromatids during yeast meiosis

Cited by 349 publications

References 44 publications

Shu1 Promotes Homolog Bias of Meiotic Recombination in Saccharomyces cerevisiae

Shu1 Promotes Homolog Bias of Meiotic Recombination in Saccharomyces cerevisiae

Red-Hed regulation: recombinase Rad51, though capable of playing the leading role, may be relegated to supporting Dmc1 in budding yeast meiosis: Figure 1.

Meiotic versus mitotic recombination: Two different routes for double‐strand break repair

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