Rad54 Protein Is Targeted to Pairing Loci by the Rad51 Nucleoprotein Filament

Mazin, Alexander V.; Bornarth, Carole J.; Solinger, Jachen A.; Heyer, Wolf Dietrich; Kowalczykowski, Stephen C.

doi:10.1016/s1097-2765(00)00057-5

Cited by 172 publications

(215 citation statements)

References 55 publications

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“…The ATPase and DNA supercoiling activities are up-regulated upon complex formation with Rad51 (Mazin et al 2000a;Van Komen et al 2000). Consistent with the results from the affinity pull-down experiments (Fig.…”

Section: Hed1 Ablates Functional Synergy Between Rad51 and Rad54supporting

confidence: 89%

Hed1 regulates Rad51-mediated recombination via a novel mechanism

Busygina¹,

Sehorn²,

Shi³

et al. 2008

Genes Dev.

104

139

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Two RecA orthologs, Rad51 and Dmc1, mediate homologous recombination in meiotic cells. During budding yeast meiosis, Hed1 coordinates the actions of Rad51 and Dmc1 by down-regulating Rad51 activity. It is thought that Hed1-dependent attenuation of Rad51 facilitates formation of crossovers that are necessary for the correct segregation of chromosomes at the first meiotic division. We purified Hed1 in order to elucidate its mechanism of action. Hed1 binds Rad51 with high affinity and specificity. We show that Hed1 does not adversely affect assembly of the Rad51 presynaptic filament, but it specifically prohibits interaction of Rad51 with Rad54, a Swi2/Snf2-like factor that is indispensable for Rad51-mediated recombination. In congruence with the biochemical results, Hed1 prevents the recruitment of Rad54 to a site-specific DNA double-strand break in vivo but has no effect on the recruitment of Rad51. These findings shed light on the function of Hed1 and, importantly, unveil a novel mechanism for the regulation of homologous recombination.[Keywords: Regulation of meiotic recombination; interhomolog crossovers; Rad51 recombinase; double-strand break repair; homologous recombination] Supplemental material is available at http://www.genesdev.org.

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Section: Hed1 Ablates Functional Synergy Between Rad51 and Rad54supporting

confidence: 89%

Hed1 regulates Rad51-mediated recombination via a novel mechanism

Busygina¹,

Sehorn²,

Shi³

et al. 2008

Genes Dev.

104

139

View full text Add to dashboard Cite

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“…The mechanism by which Rad54 protein stimulates the DNA pairing activity of Rad51 protein is different (22)(23)(24)(25)(26)(27)(28). It was found that, in contrast to other proteins that stimulate the DNA pairing activity of Rad51 protein, Rad54 protein acts at a later (synaptic) stage of DNA strand exchange (24,29).…”

mentioning

confidence: 99%

“…It was found that, in contrast to other proteins that stimulate the DNA pairing activity of Rad51 protein, Rad54 protein acts at a later (synaptic) stage of DNA strand exchange (24,29). Rad54 protein belongs to the Snf2/Swi2 group of proteins, which are involved in ATP-dependent remodeling of protein complexes, including mainly chromatin complexes (30).…”

mentioning

confidence: 99%

“…Rad54 protein was discovered to have a dsDNA topology-modifying activity, and this activity was implicated in the mechanism of stimulation of DNA strand exchange (23,27). Previously, we inferred from the analysis of joint molecule formation by Rad51 protein that Rad54 protein binds to the assembled Rad51-ssDNA nucleoprotein filament (24,31). We suggested that Rad54 protein is a component of a nucleoprotein filament that is involved in the search for DNA homology, and, in that capacity, Rad54 protein modifies the topology of dsDNA-target, thereby making it more readily accessible for DNA pairing.…”

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confidence: 99%

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A Novel Function of Rad54 Protein

Mazin¹,

Alexeev²,

Kowalczykowski³

2003

Journal of Biological Chemistry

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157

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Homologous recombination is important for the repair of double-stranded DNA breaks in all organisms. Rad51 and Rad54 proteins are two key components of the homologous recombination machinery in eukaryotes. In vitro, Rad51 protein assembles with singlestranded DNA to form the helical nucleoprotein filament that promotes DNA strand exchange, a basic step of homologous recombination. Rad54 protein interacts with this Rad51 nucleoprotein filament and stimulates its DNA pairing activity, suggesting that Rad54 protein is a component of the nucleoprotein complex involved in the DNA homology search. Here, using physical criteria, we demonstrate directly the formation of Rad54-Rad51-DNA nucleoprotein co-complexes that contain equimolar amounts of each protein. The binding of Rad54 protein significantly stabilizes the Rad51 nucleoprotein filament formed on either single-stranded DNA or double-stranded DNA. The Rad54-stabilized nucleoprotein filament is more competent in DNA strand exchange and acts over a broader range of solution conditions. Thus, the co-assembly of an interacting partner with the Rad51 nucleoprotein filament represents a novel means of stabilizing the biochemical entity central to homologous recombination, and reveals a new function of Rad54 protein.Homologous recombination plays an essential role in repair of DNA double-stranded breaks, a lethal type of DNA damage. The mechanisms of double-stranded break repair by recombination have been extensively investigated at the genetic and biochemical levels (1-6). In yeast, genetic analysis reveals a set of genes, called the RAD52 epistasis group, that are directly involved in double-stranded break repair (7). This group includes the RAD50, RAD58/MRE11, XRS2, RPA1, RAD51, RAD52, RAD54, RAD55, RAD57, and RAD59 genes. Among the proteins encoded by these genes, the Rad51 protein is the most evolutionarily conserved; its homologues are spread from bacteria to mammals (8). The Rad51 protein family members possess a unique property; they form helical nucleoprotein filaments with DNA (9 -11) and promote DNA pairing and strand transfer, a basic step of homologous recombination (11-16).More recently, it was demonstrated that Rad52 protein, Rad54 protein, and the Rad55/Rad57 heterodimer stimulate DNA pairing promoted by Rad51 protein. The mechanistic studies revealed that stimulation by Rad52 protein and the Rad55/Rad57 proteins is at the early (presynaptic) stage of DNA strand exchange. Rad52 protein enhances the ability of Rad51 protein to displace RPA from ssDNA 1 (17)(18)(19)(20), as do the Rad55/Rad57 proteins, but by a different mechanism (21).The mechanism by which Rad54 protein stimulates the DNA pairing activity of Rad51 protein is different (22-28). It was found that, in contrast to other proteins that stimulate the DNA pairing activity of Rad51 protein, Rad54 protein acts at a later (synaptic) stage of DNA strand exchange (24, 29). Rad54 protein belongs to the Snf2/Swi2 group of proteins, which are involved in ATP-dependent remodeling of protein complexes, in...

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“…Thus, Rad54 plays a major role in regulating the activities of Rad51. Reciprocally, the enzymatic activities of Rad54 -ATP hydrolysis, DNA supercoiling, DNA strand opening, DNA branch migration, and chromatin remodeling -are enhanced by Rad51 [9,[12][13][14]16,35]. In strains deleted for RAD54, ssDNA associated with a site-specific DSB at the MAT locus can synapse with its homologous donor sequence, but DNA strand invasion appears to be impaired [33,36].…”

Section: Introductionmentioning

confidence: 99%

Synergistic action of the Saccharomyces cerevisiae homologous recombination factors Rad54 and Rad51 in chromatin remodeling

et al. 2007

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Rad54, a member of the Swi2/Snf2 protein family, works in concert with the RecA-like recombinase Rad51 during the early and late stages of homologous recombination. Rad51 markedly enhances the activities of Rad54, including the induction of topological changes in DNA and the remodeling of chromatin structure. Reciprocally, Rad54 promotes Rad51-mediated DNA strand invasion with either naked or chromatinized DNA. Here, using various Saccharomyces cerevisiae rad51 and rad54 mutant proteins, mechanistic aspects of Rad54/Rad51-mediated chromatin remodeling are defined. Disruption of the Rad51-Rad54 complex leads to a marked attenuation of chromatin remodeling activity. Moreover, we present evidence that assembly of the Rad51 presynaptic filament represents an obligatory step in the enhancement of the chromatin remodeling reaction. Interestingly, we find a specific interaction of the N-terminal tail of histone H3 with Rad54 and show that the H3 tail interaction domain resides within the amino terminus of Rad54. These results suggest that Rad54-mediated chromatin remodeling coincides with DNA homology search by the Rad51 presynaptic filament and that this process is facilitated by an interaction of Rad54 with histone H3.

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Rad54 Protein Is Targeted to Pairing Loci by the Rad51 Nucleoprotein Filament

Cited by 172 publications

References 55 publications

Hed1 regulates Rad51-mediated recombination via a novel mechanism

Hed1 regulates Rad51-mediated recombination via a novel mechanism

A Novel Function of Rad54 Protein

Synergistic action of the Saccharomyces cerevisiae homologous recombination factors Rad54 and Rad51 in chromatin remodeling

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