BRCA2 is a mediator of RAD51‐ and DMC1‐facilitated homologous recombination in<i>Arabidopsis thaliana</i>

Seeliger, Katharina; Dukowic-Schulze, Stefanie; Wurz-Wildersinn, Rebecca; Pacher, Michael; Puchta, Holger

doi:10.1111/j.1469-8137.2011.03947.x

Cited by 61 publications

(73 citation statements)

References 95 publications

(229 reference statements)

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“…The primary antibodies used in this study were anti-ASY1 , anti-ZYP1 (Higgins et al, 2005), anti-RAD51 (Mercier et al, 2003), and anti-MUS81 (Higgins et al, 2008a). In the case of simultaneous staining of proteins with primary antibodies from the same species, the first primary antibody was covered with labeled Fab fragments before application of the second primary antibody (Seeliger et al, 2012). Analysis of chiasmata and statistical analysis was performed as described (Sanchez-Moran et al, 2002;Higgins et al, 2004).…”

Section: Cytological Methodsmentioning

confidence: 99%

“…Following the formation of DSBs by Spo11 (for sporulation11) (Keeney et al, 1997;Hartung and Puchta, 2000;Grelon et al, 2001;Stacey et al, 2006;Hartung et al, 2007b), recombination is initiated between homologous chromosomes by the recombinases Rad51 (for radiation sensitive51) and Dmc1 (for disrupted meiotic cDNA1) (Klimyuk and Jones, 1997;Li et al, 2004;Shinohara and Shinohara, 2004;Seeliger et al, 2012). These proteins facilitate the invasion of single-stranded ends of a DSB into homologous sequences, creating a D-loop structure.…”

Section: Introductionmentioning

confidence: 99%

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The Fanconi Anemia Ortholog FANCM Ensures Ordered Homologous Recombination in Both Somatic and Meiotic Cells in Arabidopsis

et al. 2012

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The human hereditary disease Fanconi anemia leads to severe symptoms, including developmental defects and breakdown of the hematopoietic system. It is caused by single mutations in the FANC genes, one of which encodes the DNA translocase FANCM (for Fanconi anemia complementation group M), which is required for the repair of DNA interstrand cross-links to ensure replication progression. We identified a homolog of FANCM in Arabidopsis thaliana that is not directly involved in the repair of DNA lesions but suppresses spontaneous somatic homologous recombination via a RecQ helicase (At-RECQ4A)-independent pathway. In addition, it is required for double-strand break-induced homologous recombination. The fertility of Atfancm mutant plants is compromised. Evidence suggests that during meiosis At-FANCM acts as antirecombinase to suppress ectopic recombination-dependent chromosome interactions, but this activity is antagonized by the ZMM pathway to enable the formation of interference-sensitive crossovers and chromosome synapsis. Surprisingly, mutation of At-FANCM overcomes the sterility phenotype of an At-MutS homolog4 mutant by apparently rescuing a proportion of crossover-designated recombination intermediates via a route that is likely At-MMS and UV sensitive81 dependent. However, this is insufficient to ensure the formation of an obligate crossover. Thus, At-FANCM is not only a safeguard for genome stability in somatic cells but is an important factor in the control of meiotic crossover formation.

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Section: Cytological Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The Fanconi Anemia Ortholog FANCM Ensures Ordered Homologous Recombination in Both Somatic and Meiotic Cells in Arabidopsis

et al. 2012

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“…Importantly, loss of Brca2 in plants causes chromosomal aberrations during meiosis (14). In humans, GST-pull down assays using peptide fragments of BRCA2 mapped a unique DMC1 interacting site to residues 2386-2411 (8).…”

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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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“…In Arabidopsis, defects detected by the SHR assay could be correlated with meiotic defects in RAD51C and BRCA2 mutants (Abe et al, 2005;Seeliger et al, 2012). In addition, the use of SHR lines also permitted the identification of proteins involved in processing of recombination intermediates or in postreplicative repair (Mannuss et al, 2010).…”

Section: Elucidating the Role Of Multiple Factors In Shr In Plantsmentioning

confidence: 93%

In Planta Somatic Homologous Recombination Assay Revisited: A Successful and Versatile, but Delicate Tool

Puchta

Höhn

2012

Plant Cell

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Marker-transgene-dependent lines of Arabidopsis thaliana measuring somatic homologous recombination (SHR) have been available for almost two decades. Here we discuss mechanisms of marker-gene restoration, comment on results obtained using the reporter lines, and stress how caution must be applied to avoid experimental problems or false interpretation in the use of SHR reporter lines. Although theoretically possible, we conclude that explanations other than SHR are unlikely to account for restoration of marker gene expression in the SHR lines when used with appropriate controls. We provide an overview of some of the most important achievements obtained with the SHR lines, give our view of the limitations of the system, and supply the reader with suggestions on the proper handling of the SHR lines. We are convinced that SHR lines are and will remain in the near future a valuable tool to explore the mechanism and influence of external and internal factors on genome stability and DNA repair in plants.

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BRCA2 is a mediator of RAD51‐ and DMC1‐facilitated homologous recombination inArabidopsis thaliana

Cited by 61 publications

References 95 publications

The Fanconi Anemia Ortholog FANCM Ensures Ordered Homologous Recombination in Both Somatic and Meiotic Cells in Arabidopsis

The Fanconi Anemia Ortholog FANCM Ensures Ordered Homologous Recombination in Both Somatic and Meiotic Cells in Arabidopsis

BRCA2 regulates DMC1-mediated recombination through the BRC repeats

In Planta Somatic Homologous Recombination Assay Revisited: A Successful and Versatile, but Delicate Tool

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