Rad51 Is an Accessory Factor for Dmc1-Mediated Joint Molecule Formation During Meiosis

Contreras-Shannon, Verónica; Chan, Yuen‐Ling; Grubb, Jennifer; Budke, Brian; Bishop, Douglas K.

doi:10.1126/science.1219379

Cited by 296 publications

(473 citation statements)

References 30 publications

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“…However, if nutrients reappear in the environment prior to the end of prophase, the meiotic program is aborted and cells return smoothly to the mitotic cell cycle, carrying out a regular mitotic division in which intact sister chromatids segregate regularly to opposite poles. Recently, in vivo studies have revealed a strong dependence of Dmc1 activity on Rad51 (Cloud et al, 2012;Hong et al, 2013). A DSB preferentially selects a homolog partner in both Rad51-and Dmc1-promoted recombination, in the absence or presence of the meiotic axis components Red1/Mek1/Hop1.…”

Section: Discussionmentioning

confidence: 99%

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: Discussionmentioning

confidence: 99%

Shu1 Promotes Homolog Bias of Meiotic Recombination in Saccharomyces cerevisiae

Hong

Kim

2013

Molecules and Cells

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“…Work in Saccharomyces cerevisiae and Arabidopsis has established that Rad51 is needed for Dmc1 filament formation, and one model posits that Rad51 serves to initiate a Dmc1 filament (4,21,22). In our previous work, we showed that BRC1-4 bind with high affinity to free RAD51, enhancing ssDNA assembly and limiting its nucleation on dsDNA, resulting in a stimulation of DNA strand exchange.…”

Section: Brca2 Stimulates the Dna Strand Exchange By Dmc1 Overcomingmentioning

confidence: 99%

BRCA2 regulates DMC1-mediated recombination through the BRC repeats

Martinez

Nicolai

Kim

et al. 2016

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

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In somatic cells, BRCA2 is needed for RAD51-mediated homologous recombination. The meiosis-specific DNA strand exchange protein, DMC1, promotes the formation of DNA strand invasion products (joint molecules) between homologous molecules in a fashion similar to RAD51. BRCA2 interacts directly with both human RAD51 and DMC1; in the case of RAD51, this interaction results in stimulation of RAD51-promoted DNA strand exchange. However, for DMC1, little is known regarding the basis and functional consequences of its interaction with BRCA2. Here we report that human DMC1 interacts directly with each of the BRC repeats of BRCA2, albeit most tightly with repeats 1-3 and 6-8. However, BRC1-3 bind with higher affinity to RAD51 than to DMC1, whereas BRC6-8 bind with higher affinity to DMC1, providing potential spatial organization to nascent filament formation. With the exception of BRC4, each BRC repeat stimulates joint molecule formation by DMC1. The basis for this stimulation is an enhancement of DMC1-ssDNA complex formation by the stimulatory BRC repeats. Lastly, we demonstrate that full-length BRCA2 protein stimulates DMC1-mediated DNA strand exchange between RPAssDNA complexes and duplex DNA, thus identifying BRCA2 as a mediator of DMC1 recombination function. Collectively, our results suggest unique and specialized functions for the BRC motifs of BRCA2 in promoting homologous recombination in meiotic and mitotic cells.T he breast cancer susceptibility protein 2, BRCA2, regulates RAD51-mediated homologous recombination (HR) (1-3). Both RAD51, a DNA strand exchange protein, and its meiotic counterpart, DMC1 (disrupted meiotic cDNA 1 or DNA meiotic recombinase 1), promote HR through the formation of a nucleoprotein filament on ssDNA (4). This filament finds and invades a homologous template, resulting in a DNA strand invasion product called a joint molecule or a displacement-loop (D-loop). The joint molecule provides a primer template for the new DNA synthesis required to repair the DNA double strand break (DSB).The first evidence implicating BRCA2 in meiosis came from studies in Ustilago maydis, where strains lacking the BRCA2 ortholog, Brh2, resulted in absence of meiotic products (5). Shortly thereafter, mouse BRCA2 was inferred to coordinate the activities of RAD51 and DMC1 (6). The first direct interaction between BRCA2 and DMC1 was observed in plants (7) and later in humans (8). In the plant, Arabidopsis thaliana, the interaction between Brca2 and Dmc1 was mapped to the BRC repeats (9), a highly conserved motif comprising a sequence of ∼35 amino acids that is present at least once in all BRCA2-like proteins (10). In humans, BRCA2 contains eight BRC repeats that bind with different affinities to RAD51, and they segregate into two functional classes (11). Within a BRC repeat, two motifs that bind RAD51 have been identified: one comprising the consensus sequence FxxA that mimics the oligomerization interface (12) and contacts the catalytic domain of RAD51; the other binding module comprises the alpha-helical region o...

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“…Rad51 and Dmc1 share ~45% amino acid identity (6,8,9), they assembly into structurally similar right-handed helical filaments on single-stranded DNA (ssDNA) (10)(11)(12), and they have similar, albeit not identical, biochemical properties (1,6). Despite these many commonalities, Dmc1 is the recombinase responsible for performing strand invasion during meiosis (13). In contrast, Rad51 is actively downregulated by Hed1-and Mek1-mediated inhibition of its interactions with Rad54, which is a cofactor that is required for Rad51 strand invasion activity (11,(14)(15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

Spontaneous self-segregation of Rad51 and Dmc1 DNA recombinases within mixed recombinase filaments

Crickard

Kaniecki

Kwon

et al. 2018

Journal of Biological Chemistry

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During meiosis, the two DNA recombinases Rad51 and Dmc1, form specialized presynaptic filaments that are adapted for performing recombination between homologous chromosomes. There is currently a limited understanding of how these two recombinases are organized within the meiotic presynaptic filament. Here, we used single molecule imaging to examine the properties of presynaptic complexes comprised of both Rad51 and Dmc1. We demonstrate that Rad51 and Dmc1 have an intrinsic ability to selfsegregate, even in the absence of any other recombination accessory proteins. Moreover, we found that the presence of Dmc1 stabilizes the adjacent Rad51 filaments, suggesting that crosstalk between these two recombinases may affect their biochemical properties. Based upon these findings, we describe a model for the organization of Rad51 and Dmc1 within the meiotic presynaptic complex, which is also consistent with in vivo observations, genetic findings and biochemical expectations. This model argues against the existence of extensively intermixed filaments, and we propose that Rad51 and Dmc1 have intrinsic capacities to form spatially distinct filaments, suggesting that additional recombination cofactors are not required to segregate the Rad51 and Dmc1 filaments.

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Rad51 Is an Accessory Factor for Dmc1-Mediated Joint Molecule Formation During Meiosis

Cited by 296 publications

References 30 publications

Shu1 Promotes Homolog Bias of Meiotic Recombination in Saccharomyces cerevisiae

Shu1 Promotes Homolog Bias of Meiotic Recombination in Saccharomyces cerevisiae

BRCA2 regulates DMC1-mediated recombination through the BRC repeats

Spontaneous self-segregation of Rad51 and Dmc1 DNA recombinases within mixed recombinase filaments

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