Roles of ATP binding and ATP hydrolysis in human Rad51 recombinase function

Chi, Peter; Komen, Stephen Van; Sehorn, Michael G.; Sigurdsson, Stefan; Sung, Patrick

doi:10.1016/j.dnarep.2005.11.005

Cited by 165 publications

(252 citation statements)

References 52 publications

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“…We initially used the Rad51 K133R protein, which binds ATP but is greatly attenuated for ATP hydrolysis, in the D-loop reaction. Since ATP hydrolysis prompts the dissociation of Rad51 from DNA, Rad51 K133R forms a highly stable presynaptic filament (Chi et al 2006), and its use facilitated our analyses. Coimmunoprecipitation verified that RECQL5 has the same affinity for Rad51 K133R as Rad51 (data not shown).…”

Section: Human Recql5 Physically Interacts With Rad51 and Suppresses mentioning

confidence: 99%

“…5F). Rad51 forms a functional presynaptic filament in the presence of the nonhydrolyzable ATP analog AMP-PNP (Chi et al 2006). The D-loop reaction catalyzed by Rad51 with AMP-PNP as the nucleotide cofactor is impervious to RECQL5 (data not shown), a result that further supports the conclusion that ATP hydrolysis by RECQL5 is needed for Rad51 inhibition.…”

Section: D-loop Inhibitory Action Is Dependent On Atp Hydrolysis By Rmentioning

confidence: 99%

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RECQL5/Recql5 helicase regulates homologous recombination and suppresses tumor formation via disruption of Rad51 presynaptic filaments

Hu¹,

Raynard²,

Sehorn³

et al. 2007

Genes Dev.

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287

411

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Members of the RecQ helicase family play critical roles in genome maintenance. There are five RecQ homologs in mammals, and defects in three of these (BLM, WRN, and RECQL4) give rise to cancer predisposition syndromes in humans. RECQL and RECQL5 have not been associated with a human disease. Here we show that deletion of Recql5 in mice results in cancer susceptibility. Recql5-deficient cells exhibit elevated frequencies of spontaneous DNA double-strand breaks and homologous recombination (HR) as scored using a reporter that harbors a direct repeat, and are prone to gross chromosomal rearrangements in response to replication stress. To understand how RECQL5 regulates HR, we use purified proteins to demonstrate that human RECQL5 binds the Rad51 recombinase and inhibits Rad51-mediated D-loop formation. By biochemical means and electron microscopy, we show that RECQL5 displaces Rad51 from single-stranded DNA (ssDNA) in a reaction that requires ATP hydrolysis and RPA. Together, our results identify RECQL5 as an important tumor suppressor that may act by preventing inappropriate HR events via Rad51 presynaptic filament disruption.[Keywords: Recql5 helicase; DNA repair; homologous recombination; tumor suppressor; Rad51 recombinase] Supplemental material is available at http://www.genesdev.org.

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Section: Human Recql5 Physically Interacts With Rad51 and Suppresses mentioning

confidence: 99%

Section: D-loop Inhibitory Action Is Dependent On Atp Hydrolysis By Rmentioning

confidence: 99%

RECQL5/Recql5 helicase regulates homologous recombination and suppresses tumor formation via disruption of Rad51 presynaptic filaments

Hu¹,

Raynard²,

Sehorn³

et al. 2007

Genes Dev.

Self Cite

287

411

View full text Add to dashboard Cite

show abstract

“…To promote HR, Rad51 forms a filament on the 3¢ ssDNA, which then invades and anneals to a homologous template provided by replicating sister chromatids or homologous chromatids [12]. ATPase activity of Rad51 is critical for stabilizing the catalytically active nucleoprotein filament [26]. Rad51 mutants defective for either ATP binding or ATP hydrolysis are unable to restart stalled replication forks and repair DSBs in human ESCs [27].…”

Section: Introductionmentioning

confidence: 99%

Rad51 Regulates Cell Cycle Progression by Preserving G2/M Transition in Mouse Embryonic Stem Cells

Yoon

Kim

et al. 2014

Stem Cells and Development

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Homologous recombination (HR) maintains genomic integrity against DNA replication stress and deleterious lesions, such as double-strand breaks (DSBs). Rad51 recombinase is critical for HR events that mediate the exchange of genetic information between parental chromosomes in eukaryotes. Additionally, Rad51 and HR accessory factors may facilitate replication fork progression by preventing replication fork collapse and repair DSBs that spontaneously arise during the normal cell cycle. In this study, we demonstrated a novel role for Rad51 during the cell cycle in mouse embryonic stem cells (mESCs). In mESCs, Rad51 was constitutively expressed throughout the cell cycle, and the formation of Rad51 foci increased as the cells entered S phase. Suppression of Rad51 expression caused cells to accumulate at G2/M phase and activated the DNA damage checkpoint, but it did not affect the self-renewal or differentiation capacity of mESCs. Even though Rad51 suppression significantly inhibited the proliferation rate of mESCs, Rad51 suppression did not affect the replication fork progression and speed, indicating that Rad51 repaired DNA damage and promoted DNA replication in S phase through an independent mechanism. In conclusion, Rad51 may contribute to G2/M transition in mESCs, while preserving genomic integrity in global organization of DNA replication fork.

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“…Like their prokaryotic counterparts E. coli RecA and T4 UvsX, Rad51 and Dmc1 form a filamentous structure, referred to as the presynaptic filament, on ssDNA [18,19]. Assembly of the presynaptic filament requires ATP binding by the recombinase protein [19,20], and ATP hydrolysis by the DNA-bound recombinase promotes the turnover of the presynaptic filament [21][22][23][24]. All the subsequent biochemical steps, including the capture of the duplex DNA partner and the search for homology in the engaged ssDNA and dsDNA molecules, that lead to D-loop formation occur within the confines of the presynaptic filament [17,19,20].…”

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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Roles of ATP binding and ATP hydrolysis in human Rad51 recombinase function

Cited by 165 publications

References 52 publications

RECQL5/Recql5 helicase regulates homologous recombination and suppresses tumor formation via disruption of Rad51 presynaptic filaments

RECQL5/Recql5 helicase regulates homologous recombination and suppresses tumor formation via disruption of Rad51 presynaptic filaments

Rad51 Regulates Cell Cycle Progression by Preserving G2/M Transition in Mouse Embryonic Stem Cells

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

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