Human RAD52 Captures and Holds DNA Strands, Increases DNA Flexibility, and Prevents Melting of Duplex DNA: Implications for DNA Recombination

Brouwer, Ineke; Zhang, Hongshan; Candelli, Andrea; Normanno, Davide; Peterman, Erwin J. G.; Wuite, Gijs J. L.; Modesti, Mauro

doi:10.1016/j.celrep.2017.02.068

Cited by 32 publications

(33 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This compaction can be reversed by displacement of RAD52 from the ssDNA by the addition of either 7 M urea (not shown). These observations are consistent with the ring-like structure of RAD52 and the fact that its ssDNA-binding domain follows the circular contour of this surface, which would necessitate a reduction in the apparent end-to-end distance for any bound ssDNA, as has been previously reported (51). We find some striking similarities, as well as some notable differences, between the results we report here for human RAD52 and results we have previously reported for S. cerevisiae Rad52 (65).…”

Section: Rad52 Interactions With Recombination Intermediatessupporting

confidence: 90%

“…Rad52 is highly conserved, and human RAD52 shares many biochemical traits with its yeast counterpart (42,43). Like yeast Rad52, human RAD52 also forms ring-like structures (44 -47), binds tightly to ssDNA, and promotes ssDNA annealing (47)(48)(49)(50)(51). However, human RAD52 does not have a known mediator activity (42,43), and in striking contrast to S. cerevisiae, RAD52 deletions do not produce a strong phenotype in vertebrates (42,43,52,53).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Human RAD52 interactions with replication protein A and the RAD51 presynaptic complex

Jian

Kwon

Sung

et al. 2017

Journal of Biological Chemistry

View full text Add to dashboard Cite

Rad52 is a highly conserved protein involved in the repair of DNA damage. Human RAD52 has been shown to mediate single-stranded DNA (ssDNA) and is synthetic lethal with mutations in other key recombination proteins. For this study, we used single-molecule imaging and ssDNA curtains to examine the binding interactions of human RAD52 with replication protein A (RPA)-coated ssDNA, and we monitored the fate of RAD52 during assembly of the presynaptic complex. We show that RAD52 binds tightly to the RPA-ssDNA complex and imparts an inhibitory effect on RPA turnover. We also found that during presynaptic complex assembly, most of the RPA and RAD52 was displaced from the ssDNA, but some RAD52-RPA-ssDNA complexes persisted as interspersed clusters surrounded by RAD51 filaments. Once assembled, the presence of RAD51 restricted formation of new RAD52-binding events, but additional RAD52 could bind once RAD51 dissociated from the ssDNA. Together, these results provide new insights into the behavior and dynamics of human RAD52 during presynaptic complex assembly and disassembly.

show abstract

Section: Rad52 Interactions With Recombination Intermediatessupporting

confidence: 90%

mentioning

confidence: 99%

Human RAD52 interactions with replication protein A and the RAD51 presynaptic complex

Jian

Kwon

Sung

et al. 2017

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…During RAD51--dependent GC process, RAD52 can also act on the annealing step, allowing the capture of the displaced strand by the resected second double--strand ends (Miyazaki et al, 2004;McIlwraith & West, 2008;Sugiyama et al, 2006;Brouwer et al, 2017). We show here that RAD51 blocks annealing when it is loaded on both complementary ssDNAs, which is the situation in both SSA and A--EJ.…”

Section: Discussionmentioning

confidence: 73%

Mammalian RAD51 prevents non-conservative alternative end-joining and single strand annealing through non-catalytic mechanisms

Muhammad

Chailleux

et al. 2019

Preprint

View full text Add to dashboard Cite

The selection of the DNA double-strand breaks (DSBs) repair pathway is decisive for genetic stability/instability. We proposed that it acts according to two successive steps: 1-canonical non-homologous end-joining (C-NHEJ) versus single-strand DNA (ssDNA) resection; 2-on ssDNA, gene conversion (GC) versus non-conservative singlestrand annealing (SSA) or alternative end-joining (A-EJ).Using intramolecular substrates, we systematically analysed the equilibrium between the different DSB repair pathways. We show that ablation of RAD51 stimulated both SSA and A-EJ but did not stimulate C-NHEJ, validating the two-step model. Moreover, we found that two ATP-mutant dominant-negative forms of RAD51 that stimulated non-conservative repair, failed to load into damaged chromatin, clarifying the role of ATP in RAD51-mediated HR, also. In contrast, another dominantnegative form of RAD51, which retains its DNA binding capacities, repressed SSA and A-EJ, revealing two separable functions of RAD51 i.e. GC and non-conservative repair inhibition. In vitro assays show that the binding of RAD51 on both complementary ssDNA is required to block both spontaneous and RAD52-induced strand annealing. Therefore, RAD51 represses non-conservative repair (SSA and A-EJ), by inhibiting the annealing step through ssDNA occupancy, independently of the catalytic strandexchange activity required for GC.

show abstract

“…Recently, it was reported that the DNA damage pathwaydependent regulation of Cep3 stimulates the SAC (54) and human Rad52 prevents force-induced DNA strand melting (55). In addition, some studies verified the involvement of components of the DNA damage pathway in the regulation of mitosis (35,56).…”

Section: Discussionmentioning

confidence: 97%

Rad52 phosphorylation by Ipl1 and Mps1 contributes to Mps1 kinetochore localization and spindle assembly checkpoint regulation

Lim

Huh

2017

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Rad52 is well known as a key factor in homologous recombination. Here, we report that Rad52 has functions unrelated to homologous recombination in Saccharomyces cerevisiae; it plays a role in the recruitment of Mps1 to the kinetochores and the maintenance of spindle assembly checkpoint (SAC) activity. Deletion of RAD52 causes various phenotypes related to the dysregulation of chromosome biorientation. Rad52 directly affects efficient operation of the SAC and accurate chromosome segregation. Remarkably, by using an in vitro kinase assay, we found that Rad52 is a substrate of Ipl1/Aurora and Mps1 in yeast and humans. Ipl1-dependent phosphorylation of Rad52 facilitates the kinetochore accumulation of Mps1, and Mps1-dependent phosphorylation of Rad52 is important for the accurate regulation of the SAC under spindle damage conditions. Taken together, our data provide detailed insights into the regulatory mechanism of chromosome biorientation by mitotic kinases.Rad52 | spindle assembly checkpoint | mitotic kinases | phosphorylation

show abstract

Human RAD52 Captures and Holds DNA Strands, Increases DNA Flexibility, and Prevents Melting of Duplex DNA: Implications for DNA Recombination

Cited by 32 publications

References 50 publications

Human RAD52 interactions with replication protein A and the RAD51 presynaptic complex

Human RAD52 interactions with replication protein A and the RAD51 presynaptic complex

Mammalian RAD51 prevents non-conservative alternative end-joining and single strand annealing through non-catalytic mechanisms

Rad52 phosphorylation by Ipl1 and Mps1 contributes to Mps1 kinetochore localization and spindle assembly checkpoint regulation

Contact Info

Product

Resources

About