Identification and purification of two distinct complexes containing the five RAD51 paralogs

Masson, Jean‐Yves; Tarsounas, Madalena; Stasiak, Alicja Z.; Stasiak, Andrzej; Shah, Rajvee; McIlwraith, Michael J.; Benson, Fiona E.; West, Stephen C.

doi:10.1101/gad.947001

Cited by 344 publications

(327 citation statements)

References 65 publications

(95 reference statements)

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“…Physical interactions between the five Rad51 paralogs have also been demonstrated using yeast two-hybrid experiments and coimmunoprecipitation (Schild et al, 2000;Masson et al, 2001). Rad51 paralogs form two complexes, Rad51B-Rad51C-Rad51D-Xrcc2 (BCDX2) and Rad51C-Xrcc3 (CX3), both containing Rad51C as a component (Miller et al, 2004).…”

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confidence: 99%

Arabidopsis RAD51C Gene Is Important for Homologous Recombination in Meiosis and Mitosis

et al. 2005

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Rad51 is a homolog of the bacterial RecA recombinase, and a key factor in homologous recombination in eukaryotes. Rad51 paralogs have been identified from yeast to vertebrates. Rad51 paralogs are thought to play an important role in the assembly or stabilization of Rad51 that promotes homologous pairing and strand exchange reactions. We previously characterized two RAD51 paralogous genes in Arabidopsis (Arabidopsis thaliana) named AtRAD51C and AtXRCC3, which are homologs of human RAD51C and XRCC3, respectively, and described the interaction of their products in a yeast two-hybrid system. Recent studies showed the involvement of AtXrcc3 in DNA repair and functional role in meiosis. To determine the role of RAD51C in meiotic and mitotic recombination in higher plants, we characterized a T-DNA insertion mutant of AtRAD51C. Although the atrad51C mutant grew normally during vegetative developmental stage, the mutant produced aborted siliques, and their anthers did not contain mature pollen grains. Crossing of the mutant with wild-type plants showed defective male and female gametogeneses as evidenced by lack of seed production. Furthermore, meiosis was severely disturbed in the mutant. The atrad51C mutant also showed increased sensitivity to g-irradiation and cisplatin, which are known to induce double-strand DNA breaks. The efficiency of homologous recombination in somatic cells in the mutant was markedly reduced relative to that in wild-type plants.

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confidence: 99%

Arabidopsis RAD51C Gene Is Important for Homologous Recombination in Meiosis and Mitosis

et al. 2005

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“…Yeast two-hybrid tests confirmed that human RAD51D interacts with XRCC2 and RAD51C; in addition, yeast three-hybrid system indicates that a number of different pairs of interactions between these human RAD51 paralogs may occur simultaneously [131]. Later experiments with isotope-tagged proteins suggest that RAD51B is also involved in the above complex, making a four-component complex including RAD51B, RAD51C, RAD51D, and XRCC2 (defined as BCDX2) [132]. The BCDX2 complex binds to nicks of duplex DNA and exhibits ATPase activity in vitro.…”

Section: Rad51 Paralogs In Animals and Plants: Rad51b Rad51c Rad51dmentioning

confidence: 94%

“…These results together with the observations that RAD51B [107,108], RAD51C [111], RAD51D [117], and XRCC2 [123] are required for proper formation of RAD51 foci, indicate that these proteins may function as a complex both in assembling the RAD51 complex and in the initial processing of the single-stranded DNA at the damaged sites. Another complex, with RAD51C and XRCC3, was originally detected with a yeast two-hybrid system and then with interaction of proteins expressed in baculovirusinfected insect cells [132][133][134], and then confirmed with human cells lines that express isotope-tagged XRCC3 or RAD51C proteins [135]. In addition, XRCC3 is shown to be required for stabilizing RAD51C possibly by forming dimers, and this function requires the ATPase activity, suggesting that XRCC3 may regulate the dimerization through its ATP binding and hydrolysis activity [136].…”

Section: Rad51 Paralogs In Animals and Plants: Rad51b Rad51c Rad51dmentioning

confidence: 99%

Double-stranded DNA breaks and gene functions in recombination and meiosis

Mā

2006

Cell Res

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Gene functions in recombination and meiosis 402 npg REVIEW Meiotic prophase I is a long and complex phase. Homologous recombination is an important process that occurs between homologous chromosomes during meiotic prophase I. Formation of chiasmata, which hold homologous chromosomes together until the metaphase I to anaphase I transition, is critical for proper chromosome segregation. Recent studies have suggested that the SPO11 proteins have conserved functions in a number of organisms in generating sites of double-stranded DNA breaks (DSBs) that are thought to be the starting points of homologous recombination. Processing of these sites of DSBs requires the function of RecA homologs, such as RAD51, DMC1, and others, as suggested by mutant studies; thus the failure to repair these meiotic DSBs results in abnormal chromosomal alternations, leading to disrupted meiosis. Recent discoveries on the functions of these RecA homologs have improved the understanding of the mechanisms underlying meiotic homologous recombination.Cell Research (2006) Meiosis is a highly coordinated cell division, which generates four haploid reproductive cells required for sexual reproduction. Meiosis involves one round of DNA replication and two nuclear divisions, meiosis I and meiosis II. The first nuclear division leads to the segregation of homologous chromosomes (homologs), and is unique to meiosis. During the second division, sister chromatids are separated, resulting in the formation of four haploid nuclei and followed by the meiotic cytokinesis that forms four haploid cells. Similar to mitosis, both meiosis I and meiosis II can be divided into four stages: prophase, metaphase, anaphase, and telophase.However, meiosis I differs from meiosis II as meiosis I involves homology search, pairing, interhomolog recombination, and synapsis of homologs, processes that are required for correct association and segregation of homologs. The prophase stage of meiosis I, or prophase I, has been the target of interest because of the occurrence of these major processes of chromosome interactions. During early prophase I, arms of sister chromosomes are closely associated by protein complexes called cohesins, which are removed during the metaphase I/anaphase I transition. However, the centromere regions remained associated until the segregation of sister chromatids during meiosis II. Homologous pairing is the result of interaction between homologous chromosomes relying on the homology of DNA sequences [1,2], and is considered to be a transient and non-stable association between homologous chromosomes. Pairing between homologous chromosomes facilitates the process of recombination. The recombination process confers the interexchange of genetic information between non-sister chromatids, and generates chiasmata, which ensure proper association between homologous chromosomes prior to chromosome segregation at anaphase I. Synapsis, however, is a stable association by forming synaptonemal complexes between chromosomes. Synaptonemal complexes are threeparti...

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“…The HR pathway repairs DSBs with high fidelity, as it uses one sister DNA strand as the repair template. The direct interaction of RAD51 and RAD52 proteins is an essential step in the HR pathway, and this interaction is facilitated by other important proteins such as BRCA1, BRCA2, XRCC2, and XRCC3 (10,11). In the NHEJ pathway, the heterodimer of KU70 and KU80 binds to the end of DSBs and activates the DNA-dependent protein kinase catalytic subunit-DNAPKcs.…”

Section: Introductionmentioning

confidence: 99%

Genetic polymorphisms in double-strand break DNA repair genes associated with risk of oral premalignant lesions

Yang

Lippman

Huang

et al. 2008

European Journal of Cancer

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Oral premalignant lesions (OPLs) are early genetic events en route to oral cancer. To identify individuals susceptible to OPL is critical to the prevention of oral cancer. In a case-control study consisting of 147 patients with histologically confirmed OPL and 147 matched controls, we evaluated the associations of 10 genetic variants in nine genes of the double-strand break (DSB) DNA repair pathway with OPL risk. The most notable finding was an intronic polymorphism (A17893G) of the XRCC3 gene. Compared with the homozygous wild-type AA genotype, the odds ratios [OR] (95% confidence interval [CI]) for the heterozygous AG and homozygous variant GG genotype were 0.85 (0.49-1.48) and 0.18 (0.07-0.47), respectively (P for trend=0.002). In addition, compared with the most common A-C haplotype of XRCC3 (in the order of A17893G-T241M), the G-C haplotype was associated with a significantly decreased risk of OPL (OR=0.40, 95% CI 0.23-0.68). Moreover, compared with individuals without the G-C haplotype, the ORs were 1.04 (0.56-1.95) and 0.20 (0.08-0.51) for subjects with one copy and two copies of the G-C haplotype, respectively (P for trend=0.005). Classification and regression tree (CART) analysis further revealed potential highorder gene-gene and gene-environmental interactions and categorized subjects into different risk groups according to their specific polymorphic signatures. Overall, our study provides the first epidemiological evidence supporting a connection between DSB gene variants and OPL development. Our data also suggest that the effects of high-order interactions should be taken into consideration when evaluating OPL predisposition.

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Identification and purification of two distinct complexes containing the five RAD51 paralogs

Cited by 344 publications

References 65 publications

Arabidopsis RAD51C Gene Is Important for Homologous Recombination in Meiosis and Mitosis

Arabidopsis RAD51C Gene Is Important for Homologous Recombination in Meiosis and Mitosis

Double-stranded DNA breaks and gene functions in recombination and meiosis

Genetic polymorphisms in double-strand break DNA repair genes associated with risk of oral premalignant lesions

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