Molecular Anatomy of the Recombination Mediator Function of Saccharomyces cerevisiae Rad52

Seong, Changhyun; Sehorn, Michael G.; Plate, Iben; Shi, Idina; Song, Binwei; Chi, Peter; Mortensen, Uffe Hasbro; Sung, Patrick; Krejčí, Lumír

doi:10.1074/jbc.m800763200

Cited by 55 publications

(69 citation statements)

References 58 publications

(88 reference statements)

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“…An internal region of Rad22 (amino acids 181-310) interacted with RPA, indicating that this region contained the RPA interaction domain (Fig. 4D), consistent with previous results (Seong et al 2008). The E250A-D251A double mutant of this region interacted with RPA, but the D240A-E241A mutant did not detectably do so (Fig.…”

Section: Rad22 Inhibits Dmc1 Loading Onto Rpa-coated Ssdnasupporting

confidence: 81%

Dual regulation of Dmc1-driven DNA strand exchange by Swi5–Sfr1 activation and Rad22 inhibition

Murayama¹,

Kurokawa²,

Tsutsui³

et al. 2013

Genes Dev.

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Both ubiquitously expressed Rad51 and meiosis-specific Dmc1 are required for crossover production during meiotic recombination. The budding yeast Rad52 and its fission yeast ortholog, Rad22, are “mediators;” i.e., they help load Rad51 onto ssDNA coated with replication protein A (RPA). Here we show that the Swi5–Sfr1 complex from fission yeast is both a mediator that loads Dmc1 onto ssDNA and a direct “activator” of DNA strand exchange by Dmc1. In stark contrast, Rad22 inhibits Dmc1 action by competing for its binding to RPA-coated ssDNA. Thus, Rad22 plays dual roles in regulating meiotic recombination: activating Rad51 and inhibiting Dmc1.

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Section: Rad22 Inhibits Dmc1 Loading Onto Rpa-coated Ssdnasupporting

confidence: 81%

Dual regulation of Dmc1-driven DNA strand exchange by Swi5–Sfr1 activation and Rad22 inhibition

Murayama¹,

Kurokawa²,

Tsutsui³

et al. 2013

Genes Dev.

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“…The N-terminal region of the protein harbors an oligomerization domain with DNA binding activity while the C-terminal region associates with Rad51. It had been generally assumed that the N-terminal DNA binding function was important for delivery of Rad51 to RPA-coated DNA (39), but remarkably, there is another DNA-binding domain in the C terminus that is able to promote Rad51 filament assembly and strand annealing more efficiently than the N-terminal domain (40).…”

Section: Discussionmentioning

confidence: 99%

DNA-binding Domain within the Brh2 N Terminus Is the Primary Interaction Site for Association with DNA

Zhou

Kojic

Holloman

2009

Journal of Biological Chemistry

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The C-terminal region of Brh2 (Brh2 CT ), the BRCA2 homolog in Ustilago maydis, is highly conserved and aligns with the DSS1/DNA-binding domain (DBD) of mammalian BRCA2, while the N-terminal region (Brh2 NT ) is poorly conserved and has no obvious functional domain except for the single Rad51-interacting BRC element. Paradoxically, Brh2NT , but not Brh2 CT , complements the DNA repair and recombination deficiency of the brh2 mutant. We show here that Brh2NT exhibits an unexpected DNA binding activity with properties similar to that of the full-length protein. Deletion mapping localized the region responsible for the DNA binding activity to a stretch of residues between the BRC element and the canonical DBD. A heterologous DNA-binding domain from the large subunit of replication protein A substituted for the endogenous binding region within Brh2NT in supporting DNA repair. Rad51-promoted strand invasion was stimulated by Brh2NT , but required the presence of the BRC element. The findings suggest a model in which Brh2NT serves as the principal site for association with DNA, while the Brh2 CT provides a means for regulation.The repair of damaged DNA by homologous recombination is a high fidelity and universal process for maintaining genome integrity. At the heart of it is the use of an undamaged homologous DNA sequence as a template to direct repair of damaged regions. In eukaryotes, Rad51 plays the key role of promoting the search for the homologous sequence (1). Rad51 assembles into a complex with single-stranded (ss) 2 DNA that has been revealed after some processing of the damaged duplex to form a nucleoprotein filament, which is the active element for promoting homologous pairing and DNA strand exchange. BRCA2 is an important regulator of Rad51 activity (2, 3). Investigations using peptides or synthetic fusion proteins designed to model component elements of human BRCA2 suggest both positive and negative roles for BRCA2 in organizing Rad51 filaments (4 -8).Biochemical studies with Brh2, the BRCA2 homolog from the fungus Ustilago maydis, have demonstrated that it mediates Rad51 assembly on a protruding single-strand DNA tail (9). Interaction of Rad51 with Brh2 is through the single BRC element located in the N-terminal region of the protein (10). The BRC element comprises a sequence of about 30 amino acids (aa) that mimics a critical determinant of the Rad51 polymerization interface (11,12). Rad51 also interacts with Brh2 through a second, poorly defined locus, the CRE, located at the extreme C terminus of Brh2 (13). These N-terminal and C-terminal elements communicate with each other in some as yet unknown way to support proper Rad51 filament assembly. Such communication requires an interplay that is coordinated by Dss1 (13), a small versatile acidic protein that interacts with Brh2 in a region corresponding to a segment of the DSS1/DNAbinding domain (DBD) in mammalian BRCA2 (14). The mammalian BRCA2 DBD domain consists of a tandem array of three oligonucleotide/oligosaccharide-binding (OB) folds and a helix-r...

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“…The Rad51 and Rad52 proteins were purified as described previously (47,59). The RSC complex was purified from yeast cells that chromosomally harbor tandem affinity purification (TAP)-tagged Rsc2 as described previously (69).…”

Section: Methodsmentioning

confidence: 99%

RSC Facilitates Rad59-Dependent Homologous Recombination between Sister Chromatids by Promoting Cohesin Loading at DNA Double-Strand Breaks

Oum

Seong

Kwon

et al. 2011

Molecular and Cellular Biology

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Homologous recombination repairs DNA double-strand breaks by searching for, invading, and copying information from a homologous template, typically the homologous chromosome or sister chromatid. Tight wrapping of DNA around histone octamers, however, impedes access of repair proteins to DNA damage. To facilitate DNA repair, modifications of histones and energy-dependent remodeling of chromatin are required, but the precise mechanisms by which chromatin modification and remodeling enzymes contribute to homologous DNA repair are unknown. Here we have systematically assessed the role of budding yeast RSC (remodel structure of chromatin), an abundant, ATP-dependent chromatin-remodeling complex, in the cellular response to spontaneous and induced DNA damage. RSC physically interacts with the recombination protein Rad59 and functions in homologous recombination. Multiple recombination assays revealed that RSC is uniquely required for recombination between sister chromatids by virtue of its ability to recruit cohesin at DNA breaks and thereby promoting sister chromatid cohesion. This study provides molecular insights into how chromatin remodeling contributes to DNA repair and maintenance of chromatin fidelity in the face of DNA damage.

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Molecular Anatomy of the Recombination Mediator Function of Saccharomyces cerevisiae Rad52

Cited by 55 publications

References 58 publications

Dual regulation of Dmc1-driven DNA strand exchange by Swi5–Sfr1 activation and Rad22 inhibition

Dual regulation of Dmc1-driven DNA strand exchange by Swi5–Sfr1 activation and Rad22 inhibition

DNA-binding Domain within the Brh2 N Terminus Is the Primary Interaction Site for Association with DNA

RSC Facilitates Rad59-Dependent Homologous Recombination between Sister Chromatids by Promoting Cohesin Loading at DNA Double-Strand Breaks

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