Recombination activities of HsDmc1 protein, the meiotic human homolog of RecA protein

Li, Zhufang; Golub, Efim I.; Gupta, Ravindra C.; Radding, Charles M.

doi:10.1073/pnas.94.21.11221

Cited by 130 publications

(125 citation statements)

References 41 publications

Supporting

Mentioning

116

Contrasting

Order By: Relevance

“…Thus, a large number of sequencing reads containing long ITRs can be found in the sequencing libraries prepared from meiotic cells. These reads were derived from DNA fragments that preferentially interact with DMC1 and RAD51, both known to bind ssDNA (Sung 1994;Li et al 1997). These data are thus consistent with ssDNA recovery via a hairpin-mediated mechanism.…”

Section: Resultssupporting

confidence: 70%

See 1 more Smart Citation

Sensitive mapping of recombination hotspots using sequencing-based detection of ssDNA

Khil¹,

Smagulova²,

Brick³

et al. 2012

Genome Res.

121

139

View full text Add to dashboard Cite

Meiotic DNA double-stranded breaks (DSBs) initiate genetic recombination in discrete areas of the genome called recombination hotspots. DSBs can be directly mapped using chromatin immunoprecipitation followed by sequencing (ChIPseq). Nevertheless, the genome-wide mapping of recombination hotspots in mammals is still a challenge due to the low frequency of recombination, high heterogeneity of the germ cell population, and the relatively low efficiency of ChIP. To overcome these limitations we have developed a novel method-single-stranded DNA (ssDNA) sequencing (SSDS)-that specifically detects protein-bound single-stranded DNA at DSB ends. SSDS comprises a computational framework for the specific detection of ssDNA-derived reads in a sequencing library and a new library preparation procedure for the enrichment of fragments originating from ssDNA. The use of our technique reduces the nonspecific double-stranded DNA (dsDNA) background >10-fold. Our method can be extended to other systems where the identification of ssDNA or DSBs is desired.

show abstract

Section: Resultssupporting

confidence: 70%

“…One can imagine further applications where the ability of our approach to distinguish single-stranded and double-stranded substrates would be beneficial. It may, for example, allow us to differentiate ssDNA and dsDNA targets of proteins that can bind both ssDNA and dsDNA, such as RAD51, DMC1, or Smc5 (Sung 1994;Li et al 1997;Roy et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Sensitive mapping of recombination hotspots using sequencing-based detection of ssDNA

Khil¹,

Smagulova²,

Brick³

et al. 2012

Genome Res.

121

139

View full text Add to dashboard Cite

show abstract

“…Dmc1 was also proposed to be a structural and functional relative of RecA based on mutant and sequence analysis (Bishop et al 1992;Story et al 1993). Biochemical demonstration of recombinase activity for Rad51 and Dmc1 supports the idea that each protein promotes strand exchange by a mechanism related to that of RecA (Sung 1994;Baumann et al 1996;Li et al 1997;Masson et al 1999;Hong et al 2001). Although similar in several respects, electron microscopy (EM) analysis showed that Dmc1 and Rad51 differ in that Dmc1 has a strong tendency to form tori and can bind DNA in the toroid form Passy et al 1999;Kinebuchi et al 2004).…”

mentioning

confidence: 95%

Red-Hed regulation: recombinase Rad51, though capable of playing the leading role, may be relegated to supporting Dmc1 in budding yeast meiosis: Figure 1.

Sheridan

Bishop²

2006

Genes Dev.

View full text Add to dashboard Cite

DNA strand exchange is the central process of homologous recombination. In budding yeast, this reaction is catalyzed by the recombinases Rad51 and Dmc1. Rad51 is responsible for recombinational repair of DNA damage during mitosis and is also important during meiotic recombination. Dmc1 is only expressed during meiosis and is required for meiotic recombination. The discovery that these two different recombinases are needed for normal levels of meiotic recombination in budding yeast raised the question of how the functions of these proteins relate to one another. The paper by Tsubouchi and Roeder (2006) in this issue of Genes & Development represents important progress toward what is apparently a rather complex answer. Tsubouchi and Roeder (2006) report discovery of a novel meiosis-specific protein, Hed1, which appears to inhibit Rad51 when Dmc1 is absent. The authors show mutation of the HED1 gene allows cells to bypass the meiotic arrest caused by a dmc1 mutation and resolve meiosis-specific double-strand breaks (DSBs) using Rad51. A hed1 mutation can also suppress the defects caused by mutation of HOP2, which codes for a Dmc1 accessory factor. Two-hybrid experiments show that Hed1 and Rad51 can interact directly and immuno-staining shows that subnuclear foci formed by Hed1 and Rad51 colocalize. Together these results imply that Hed1's influence on Rad51 activity is direct. Furthermore, expression of Hed1 in mitotic cells provided evidence that Hed1 expression can be sufficient to inhibit Rad51-dependent repair of mitotic DNA damage.A reasonable interpretation of these results is that the meiotic recombination machinery of budding yeast is regulated such that Dmc1 carries out most strand exchange. Tsubouchi and Roeder's (2006) results also add to evidence indicating that Rad51 is capable of replacing Dmc1's function under certain circumstances. Here, we place these new findings in the context of previous observations and present a model to explain how Rad51 and Dmc1 may cooperate to promote interhomolog recombination in budding yeast, while at the same time accounting for the observed functional redundancy. We further speculate as to how the functional relationship of Rad51 and Dmc1 may have evolved.The notion that Rad51 activity is blocked during meiosis is not new. Previous work showed that Dmc1-independent recombination is inhibited by proteins associated with axial elements (Schwacha and Kleckner 1997;Xu et al. 1997;Bishop et al. 1999;Niu et al. 2005) proteinaceous structures that organize pairs of sister chromatids into two parallel sets of loops by binding at their base. The assembly of the synaptonemal complex brings an axial element with its pair of sisters into close parallel alignment with another axial element holding the homologous sister pair. Three interacting axis-associated proteins-Red1, Hop1, and Mek1-regulate recombination by suppressing Dmc1-independent recombination and, in the case of Red1 and Hop1, by enhancing the efficiency of DSB formation in a locus-specific manner (Rockmill and Roeder...

show abstract

“…The molecular activities of RAD51 and DMC1 proteins have been studied in depth in vitro. Both proteins form helical structures on ssDNA and mediate the invasion of those so-called presynaptic filaments into homologous duplex DNA to form displacement loops (Shinohara et al, 1992;Ogawa et al, 1993;Benson et al, 1994;Sung, 1994;Baumann et al, 1996;Li et al, 1997;Masson et al, 1999;Hong et al, 2001;Sehorn et al, 2004;Yu and Egelman, 2010). Although similar in structure, some differences between the two recombinases may exist.…”

Section: Introductionmentioning

confidence: 99%

The Recombinases DMC1 and RAD51 Are Functionally and Spatially Separated during Meiosis in Arabidopsis

et al. 2012

View full text Add to dashboard Cite

Meiosis ensures the reduction of the genome before the formation of generative cells and promotes the exchange of genetic information between homologous chromosomes by recombination. Essential for these events are programmed DNA double strand breaks (DSBs) providing single-stranded DNA overhangs after their processing. These overhangs, together with the RADiation sensitive51 (RAD51) and DMC1 Disrupted Meiotic cDNA1 (DMC1) recombinases, mediate the search for homologous sequences. Current models propose that the two ends flanking a meiotic DSB have different fates during DNA repair, but the molecular details remained elusive. Here we present evidence, obtained in the model plant Arabidopsis thaliana, that the two recombinases, RAD51 and DMC1, localize to opposite sides of a meiotic DSB. We further demonstrate that the ATR kinase is involved in regulating DMC1 deposition at meiotic DSB sites, and that its elimination allows DMC1-mediated meiotic DSB repair even in the absence of RAD51. DMC1's ability to promote interhomolog DSB repair is not a property of the protein itself but the consequence of an ASYNAPTIC1 (Hop1)-mediated impediment for intersister repair. Taken together, these results demonstrate that DMC1 functions independently and spatially separated from RAD51 during meiosis and that ATR is an integral part of the regular meiotic program.

show abstract

Recombination activities of HsDmc1 protein, the meiotic human homolog of RecA protein

Cited by 130 publications

References 41 publications

Sensitive mapping of recombination hotspots using sequencing-based detection of ssDNA

Sensitive mapping of recombination hotspots using sequencing-based detection of ssDNA

Red-Hed regulation: recombinase Rad51, though capable of playing the leading role, may be relegated to supporting Dmc1 in budding yeast meiosis: Figure 1.

The Recombinases DMC1 and RAD51 Are Functionally and Spatially Separated during Meiosis in Arabidopsis

Contact Info

Product

Resources

About