Stimulation of Dmc1-mediated DNA strand exchange by the human Rad54B protein

Sarai, Naoyuki; Kagawa, Wataru; Kinebuchi, Takashi; Kagawa, Ako; Tanaka, Kozo; Miyagawa, Kiyoshi; Ikawa, Shiro; Shibata, Takehiko; Kurumizaka, Hitoshi; Yokoyama, Shigeyuki

doi:10.1093/nar/gkl562

Cited by 20 publications

(22 citation statements)

References 60 publications

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“…The stimulation of the complex formation between S100A11 and Rad54B is obviously time dependent, with the highest number of S100A11-Rad54B complexes after approx. 3 h. These kinetics are also similar to the temporal stimulation of the Dmc1 recombinase-mediated DNA strand exchange activity by Rad54B (Sarai et al, 2006). As reported, after transfer of S100A11 into the nucleus, an induction of the CDK inhibitor p21 WAF1/CIP1 is initiated (Sakaguchi et al, 2003).…”

Section: Discussionsupporting

confidence: 82%

Rad54B Targeting to DNA Double-Strand Break Repair Sites Requires Complex Formation with S100A11

Murzik

Hemmerich

Weidtkamp‐Peters

et al. 2008

MBoC

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S100A11 is involved in a variety of intracellular activities such as growth regulation and differentiation. To gain more insight into the physiological role of endogenously expressed S100A11, we used a proteomic approach to detect and identify interacting proteins in vivo. Hereby, we were able to detect a specific interaction between S100A11 and Rad54B, which could be confirmed under in vivo conditions. Rad54B, a DNA-dependent ATPase, is described to be involved in recombinational repair of DNA damage, including DNA double-strand breaks (DSBs). Treatment with bleomycin, which induces DSBs, revealed an increase in the degree of colocalization between S100A11 and Rad54B. Furthermore, S100A11/ Rad54B foci are spatially associated with sites of DNA DSB repair. Furthermore, while the expression of p21 WAF1/CIP1 was increased in parallel with DNA damage, its protein level was drastically down-regulated in damaged cells after S100A11 knockdown. Down-regulation of S100A11 by RNA interference also abolished Rad54B targeting to DSBs. Additionally, S100A11 down-regulated HaCaT cells showed a restricted proliferation capacity and an increase of the apoptotic cell fraction. These observations suggest that S100A11 targets Rad54B to sites of DNA DSB repair sites and identify a novel function for S100A11 in p21-based regulation of cell cycle. INTRODUCTIONProteins may exist in several complexes in a spatial and temporal manner to accomplish distinct functions. Analyses of the interacting partners will provide a strong insight into the physiological role of a particular factor. Therefore, it is essential to identify ideally all interacting partners of proteins in vivo to precisely be able to define their biological function. The investigation of protein complexes of solely endogenously expressed proteins avoid the tendency to detect false positive protein-protein interactions of examinations performed in vitro. A multitude of proteins are involved in both the detection and the repair of DNA damages. It is conceivable that some protein complexes involved in these processes are not yet discovered. A severe form of DNA damage that threaten the integrity of the genome are DNA double-strand breaks (DSBs). DSBs can lead to cell cycle arrest or illegitimate DNA rearrangements that can contribute to cell dysfunction, cell death, or carcinogenesis (Hoeijmakers, 2001). Homologous recombination is a major DNA repair pathway by which DSBs are repaired (Lettier et al., 2006).For the identification of specific interacting proteins of S100A11 (S100C, calgizzarin) we used in the present study a proteomic approach comprising mass spectrometry and immunological techniques. S100A11 belongs to the group of S100 proteins that are considered as multitasking proteins involved in several biological processes such as the Ca 2ϩ signaling network, cell growth and motility, cell cycle progression, transcription, and cell differentiation (Schafer and Heizmann, 1996;Donato, 2001;Eckert et al., 2004). It has been proposed that the S100 proteins are involved in...

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Section: Discussionsupporting

confidence: 82%

Rad54B Targeting to DNA Double-Strand Break Repair Sites Requires Complex Formation with S100A11

Murzik

Hemmerich

Weidtkamp‐Peters

et al. 2008

MBoC

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“…hRAD54B interacts with human RAD51 and DMC1, and enhances the D-loop formation of both proteins [64, 169, 170]. Further work is needed to understand the functional specification between hRAD54 and hRAD54B in vertebrates.…”

Section: Functions Of the Rad54 Paralogs Scrdh54 And Hrad54bmentioning

confidence: 99%

Functions of the Snf2/Swi2 family Rad54 motor protein in homologous recombination

Ceballos

Heyer

2011

Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanism

130

141

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Homologous recombination is a central pathway to maintain genomic stability and is involved in the repair of DNA damage and replication fork support, as well as accurate chromosome segregation during meiosis. Rad54 is a dsDNA-dependent ATPase of the Snf2/Swi2 family of SF2 helicases, although Rad54 lacks classical helicase activity and cannot carry out the strand displacement reactions typical for DNA helicases. Rad54 is a potent and processive motor protein that translocates on dsDNA, potentially executing several functions in recombinational DNA repair. Rad54 acts in concert with Rad51, the central protein of recombination that performs the key reactions of homology search and DNA strand invasion. Here, we will review the role of the Rad54 protein in homologous recombination with an emphasis on mechanistic studies with the yeast and human enzymes. We will discuss how these results relate to in vivo functions of Rad54 during homologous recombination in somatic cells and during meiosis.

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“…For RecA (Kowalczykowski et al 1994;Papavinasasundaram et al 1997;Voloshin et al 2001) and Rad51 (Sung et al 2003;Krogh and Symington 2004) a number of well-characterized factors have been described. For Dmc1, however, the detailed biochemical mechanisms of its accessory proteins are unknown (Chen et al 2004;Hayase et al 2004;Sehorn et al 2004;Petukhova et al 2005;Haruta et al 2006;Pezza et al 2006;Sarai et al 2006). Both genetics and biochemical studies have demonstrated the functional interaction between Dmc1 and Hop2/Mnd1 proteins.…”

Section: A Presynaptic and Synaptic Role For Hop2/mnd1 In Meiotic Recmentioning

confidence: 99%

Hop2/Mnd1 acts on two critical steps in Dmc1-promoted homologous pairing

et al. 2007

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Meiotic recombination between homologous chromosomes ensures their proper segregation at the first division of meiosis and is the main force shaping genetic variation of genomes. The HOP2 and MND1 genes are essential for this recombination: Their disruption results in severe defects in homologous chromosome synapsis and an early-stage failure in meiotic recombination. The mouse Hop2 and Mnd1 proteins form a stable heterodimer (Hop2/Mnd1) that greatly enhances Dmc1-mediated strand invasion. In order to elucidate the mechanism by which Hop2/Mnd1 stimulates Dmc1, we identify several intermediate steps in the homologous pairing reaction promoted by Dmc1. We show that Hop2/Mnd1 greatly stimulates Dmc1 to promote synaptic complex formation on long duplex DNAs, a step previously revealed only for bacterial homologous recombinases. This synaptic alignment is a consequence of the ability of Hop2/Mnd1 to (1) stabilize Dmc1-single-stranded DNA (ssDNA) nucleoprotein complexes, and (2) facilitate the conjoining of DNA molecules through the capture of double-stranded DNA by the Dmc1-ssDNA nucleoprotein filament. To our knowledge, Hop2/Mnd1 is the first homologous recombinase accessory protein that acts on these two separate and critical steps in mammalian meiotic recombination.[Keywords: DNA repair; Dmc1 recombinase; homologous recombination; strand invasion; synaptic complex] Supplemental material is available at http://www.genesdev.org. Homologous recombination serves a critical function in the repair of DNA double-strand breaks (DSBs) and in the proper segregation of homologous chromosomes in meiosis (Kleckner 1996;Roeder 1997). Failure to establish a physical connection through chiasmata causes missegregation of chromosomes at prophase I and results in meiotic cell apoptosis or aneuploid gametes. The Dmc1 recombinase, a eukaryotic homolog of the bacterial RecA protein, is expressed exclusively in meiotic cells and is a major player in meiotic homologous recombination. It promotes the search for homology and catalyzes the invasion of a single-stranded end generated by the 5Ј resection of DSBs introduced by Spo11 into a homologous unbroken double-stranded DNA (dsDNA) to form joint molecules through strand invasion (D-loop formation) (Li et al. 1997;Masson et al. 1999;Hong et al. 2001;Masson and West 2001;Neale and Keeney 2006). The interaction between Hop2, Mnd1, and Dmc1 and/or Rad51, the ubiquitously expressed eukaryotic homolog of RecA, is crucial for the progression of meiotic homologous recombination. Biochemical studies have shown that the Hop2/Mnd1 complex physically interacts with and stimulates Dmc1 and Rad51 strand invasion activity (Chen et al. 2004;Petukhova et al. 2005;Pezza et al. 2006). The cooperation between Dmc1/Rad51 and Hop2/ Mnd1 is likely to be crucial in vivo, since without Hop2 and/or Mnd1, in yeast (Leu et al. 1998;Gerton and DeRisi 2002; Roeder 2002, 2003;Zierhut et al. 2004;Henry et al. 2006), Arabidopsis thaliana (Domenichini et al. 2006;Kerzendorfer et al. 2006;Panoli et al. 2006), and mouse (Pe...

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Stimulation of Dmc1-mediated DNA strand exchange by the human Rad54B protein

Cited by 20 publications

References 60 publications

Rad54B Targeting to DNA Double-Strand Break Repair Sites Requires Complex Formation with S100A11

Rad54B Targeting to DNA Double-Strand Break Repair Sites Requires Complex Formation with S100A11

Functions of the Snf2/Swi2 family Rad54 motor protein in homologous recombination

Hop2/Mnd1 acts on two critical steps in Dmc1-promoted homologous pairing

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