Analysis of the impact of the absence of RAD51 strand exchange activity in Arabidopsis meiosis

Singh, Gunjita; Ines, Olivier Da; Gallego, María; White, Charles I.

doi:10.1371/journal.pone.0183006

Cited by 22 publications

(27 citation statements)

References 70 publications

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“…It also, however, invites speculation concerning whether it is necessary to invoke such a downregulation to explain numbers of CO vs non-CO recombination events in plants, and very likely in vertebrates. Previous work has shown that DMC1 is capable of catalysing repair of all meiotic DSB in Arabidopsis in strand-invasion mutants of RAD51 [50, 90], or as shown here, by blocking RAD51 activity through the absence of RAD54. In both of these contexts, no evidence of alteration of numbers nor distribution of meiotic recombination has been found.…”

Section: Discussionsupporting

confidence: 66%

“…Interestingly, no apparent Hed1 or Mek1 orthologues have been identified in higher eukaryotes and in particular in plants. Several reports suggest that RAD51 is also down-regulated in Arabidopsis meiosis [50, 82, 83, 87, 90, 91], but the evidence for this remains indirect. Thus, whether RAD51 strand exchange activity is down-regulated during meiosis in higher organisms and if so, how this is achieved, is not clear.…”

Section: Discussionmentioning

confidence: 99%

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RAD54 is essential for RAD51-mediated repair of meiotic DSB in Arabidopsis

Sanchez-Rebato

Bouatta

Gallego

et al. 2020

Preprint

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22An essential component of the homologous recombination machinery in eukaryotes, 23 the RAD54 protein is a member of the SWI2/SNF2 family of helicases with dsDNA-dependent 24 ATPase, DNA translocase, DNA supercoiling and chromatin remodelling activities. It is a motor 25 protein that translocates along dsDNA and performs multiple functions in homologous 26 recombination. In particular, RAD54 is an essential cofactor for regulating RAD51 activity. It 27 stabilizes the RAD51 nucleofilament, remodels nucleosomes, and stimulates homology search 28 and strand invasion activity of RAD51. Accordingly, deletion of RAD54 has dramatic 29 consequences on DNA damage repair in mitotic cells. In contrast, its role in meiotic 30 recombination is less clear. 31RAD54 is essential for meiotic recombination in Drosophila and C. elegans, but plays 32 minor roles in yeast and mammals. We present here characterization of the roles of RAD54 in 33 meiotic recombination in the model plant Arabidopsis thaliana. Absence of RAD54 has no 34 detectable effect on meiotic recombination in otherwise wild-type plants but RAD54 becomes 35 essential for meiotic DSB repair in absence of DMC1. In Arabidopsis, dmc1 mutants have an 36 achiasmate meiosis, in which RAD51 repairs meiotic DSBs. Absence of RAD54 in dmc1 37 mutants leads to meiotic chromosomal fragmentation. The action of RAD54 in meiotic RAD51 38 activity is thus downstream of the role of RAD51 in supporting the activity of DMC1. Equivalent 39 analyses show no effect on meiosis of combining dmc1 with the mutants of the RAD51-40 mediators RAD51B, RAD51D and XRCC2. 41RAD54 is thus required for repair of meiotic DSBs by RAD51 and the absence of 42 meiotic phenotype in rad54 plants is a consequence of RAD51 playing a RAD54-independent 43 supporting role to DMC1 in meiotic recombination. 44 45 46 3 47 Author Summary48 Homologous recombination is a universal pathway which repairs broken DNA molecules 49 through the use of homologous DNA templates. It is both essential for maintenance of genome 50 stability and for the generation of genetic diversity through sexual reproduction. A central step 51 of the homologous recombination process is the search for and invasion of a homologous 52 intact DNA sequence that will be used as template. This key step is catalysed by the RAD51 53 recombinase in somatic cells and RAD51 and DMC1 in meiotic cells, assisted by a number of 54 associated factors. Among these, the chromatin-remodelling protein RAD54 is a required 55 cofactor for RAD51 in mitotic cells. Understanding of its role during meiotic recombination 56 however remains elusive. We show here that RAD54 is required for repair of meiotic double 57 strand breaks by RAD51 in the plant Arabidopsis thaliana, and this function is downstream of 58 the meiotic role of RAD51 in supporting the activity of DMC1. These results provide new 59 insights into the regulation of the central step of homologous recombination in plants and very 60 probably also other multicellular eukaryotes. 61 4 62 Introduction 63 Homolo...

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Section: Discussionsupporting

confidence: 66%

Section: Discussionmentioning

confidence: 99%

RAD54 is essential for RAD51-mediated repair of meiotic DSB in Arabidopsis

Sanchez-Rebato

Bouatta

Gallego

et al. 2020

Preprint

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“…RAD51 performs strand exchange in meiosis in Caenorhabditis elegans , which lacks DMC1 ( Woglar and Villeneuve, 2018 ), and also in Schizosaccharomyces pombe ( Murayama et al., 2008 ) in combination with DMC1. RAD51 is necessary for meiosis in Saccharomyces cerevisiae and Arabidopsis thaliana ( Cloud et al., 2012 ; Da Ines et al., 2013 ), but its ability to perform strand exchange is dispensable ( Bishop, 2012 ; Cloud et al., 2012 ; Da Ines et al., 2013 ; Singh et al., 2017 ). Instead, it has a role in promoting the formation and function of DMC1 filaments ( Bishop, 2012 ; Chan et al., 2019 ).…”

Section: Introductionmentioning

confidence: 99%

The Configuration of RPA, RAD51, and DMC1 Binding in Meiosis Reveals the Nature of Critical Recombination Intermediates

et al. 2020

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Summary Meiotic recombination proceeds via binding of RPA, RAD51, and DMC1 to single-stranded DNA (ssDNA) substrates created after formation of programmed DNA double-strand breaks. Here we report high-resolution in vivo maps of RPA and RAD51 in meiosis, mapping their binding locations and lifespans to individual homologous chromosomes using a genetically engineered hybrid mouse. Together with high-resolution microscopy and DMC1 binding maps, we show that DMC1 and RAD51 have distinct spatial localization on ssDNA: DMC1 binds near the break site, and RAD51 binds away from it. We characterize inter-homolog recombination intermediates bound by RPA in vivo , with properties expected for the critical displacement loop (D-loop) intermediates. These data support the hypothesis that DMC1, not RAD51, performs strand exchange in mammalian meiosis. RPA-bound D-loops can be resolved as crossovers or non-crossovers, but crossover-destined D-loops may have longer lifespans. D-loops resemble crossover gene conversions in size, but their extent is similar in both repair pathways.

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“…It is suggested that DMC1 promotes only the CO recombination with the homologous chromosome, which is unique to meiosis, and RAD51 plays its role mainly in sister chromatid exchange or the NCO recombination (Shinohara and Shinohara, 2004; Neale and Keeney, 2006). However, a recent work has shown that in the case of absence of the RAD51-mediated strand exchange activity, the DMC1 activity is sufficient to repair all DSBs during meiosis into both CO and NCO products and it does not affect meiotic crossing-over rates or patterns (Cloud et al, 2012; Da Ines et al, 2013; Singh et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

The dmc1 Mutant Allows an Insight Into the DNA Double-Strand Break Repair During Meiosis in Barley (Hordeum vulgare L.)

Szurman-Zubrzycka

Baran

Stolarek-Januszkiewicz

et al. 2019

Front. Plant Sci.

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Meiosis is a process of essential importance for sexual reproduction, as it leads to production of gametes. The recombination event (crossing-over) generates genetic variation by introducing new combination of alleles. The first step of crossing-over is introduction of a targeted double-strand break (DSB) in DNA. DMC1 (Disrupted Meiotic cDNA1) is a recombinase that is specific only for cells undergoing meiosis and takes part in repair of such DSBs by searching and invading homologous sequences that are subsequently used as a template for the repair process. Although role of the DMC1 gene has been validated in Arabidopsis thaliana , a functional analysis of its homolog in barley, a crop species of significant importance in agriculture, has never been performed. Here, we describe the identification of barley mutants carrying substitutions in the HvDMC1 gene. We performed mutational screening using TILLING (Targeting Induced Local Lesions IN Genomes) strategy and the barley TILLING population, Hor TILLUS, developed after double-treatment of spring barley cultivar ‘Sebastian’ with sodium azide and N -methyl- N -nitrosourea. One of the identified alleles, dmc1.c , was found independently in two different M 2 plants. The G2571A mutation identified in this allele leads to a substitution of the highly conserved amino acid (arginine-183 to lysine) in the DMC1 protein sequence. Two mutant lines carrying the same dmc1.c allele show similar disturbances during meiosis. The chromosomal aberrations included anaphase bridges and chromosome fragments in anaphase/telophase I and anaphase/telophase II, as well as micronuclei in tetrads. Moreover, atypical tetrads containing three or five cells were observed. A highly increased frequency of all chromosome aberrations during meiosis have been observed in the dmc1.c mutants compared to parental variety. The results indicated that DMC1 is required for the DSB repair, crossing-over and proper chromosome disjunction during meiosis in barley.

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Analysis of the impact of the absence of RAD51 strand exchange activity in Arabidopsis meiosis

Cited by 22 publications

References 70 publications

RAD54 is essential for RAD51-mediated repair of meiotic DSB in Arabidopsis

RAD54 is essential for RAD51-mediated repair of meiotic DSB in Arabidopsis

The Configuration of RPA, RAD51, and DMC1 Binding in Meiosis Reveals the Nature of Critical Recombination Intermediates

The dmc1 Mutant Allows an Insight Into the DNA Double-Strand Break Repair During Meiosis in Barley (Hordeum vulgare L.)

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