Distinct pathways of homologous recombination controlled by the SWS1-SWSAP1-SPIDR complex

Abstract

Homology-directed repair (HDR), a critical DNA repair pathway in mammalian cells, is complex, leading to multiple outcomes with different impacts on genomic integrity. However, the factors that control these different outcomes are often not well understood. Here we show that SWS1-SWSAP1-SPIDR controls distinct types of HDR. Despite their requirement for stable assembly of RAD51 recombinase at DNA damage sites, these proteins are not essential for intrachromosomal HDR, providing insight into why patients and mice with mutations are viable.However, SWS1-SWSAP1-SPIDR is critical for inter-homolog HDR, the first mitotic factor identified specifically for this function. Furthermore, SWS1-SWSAP1-SPIDR drives the high level of sister-chromatid exchange, promotes long-range loss of heterozygosity often involved with cancer initiation, and impels the poor growth of BLM helicase-deficient cells. The relevance of these genetic interactions is evident as SWSAP1 loss prolongs Blm-mutant embryo survival, suggesting a possible druggable target for the treatment of Bloom syndrome. MainDouble-strand breaks (DSBs) are among the most dangerous DNA lesions that can arise in cells and, if not repaired correctly, can lead to genomic instability and tumorigenesis 1 . Homologous recombination, also known as homology-directed repair (HDR), is the predominant pathway to repair DSBs in an error-free manner. The RAD51 recombinase plays a central role, whereby it forms filaments on single-stranded DNA at resected DNA ends through the activity of mediator proteins like BRCA2 2 and can be stabilized and/or remodeled by RAD51 paralogs, as shown for the yeast and worm proteins 3,4 . RAD51 nucleoprotein filaments subsequently invade a homologous template, typically the sister chromatid, to form a displacement loop (D loop), followed by repair synthesis. DNA helicases can unwind these D loops to promote HDR by the synthesis-dependent strand annealing pathway to result in noncrossovers. Alternatively, D loops that capture the other DNA end can mature into double Holliday junctions, which can be dissolved by BLM, a RECQ helicase deficient in individuals with Bloom syndrome, or resolved by structure-specific nucleases to form crossovers 5,6 . What controls the decision points for these various pathways is not well understood.SWS1-SWSAP1 is a recently identified complex that is critically important for HDR during meiosis in the mouse, promoting the stable assembly of both RAD51 and DMC1 nucleoprotein filaments at resected DNA ends in spermatocytes 7 . As a result, meiotic chromosomes in Sws1 and Swsap1 mutant mice often fail to synapse properly and mice are sterile. SWS1 homologs are readily

DNA Repair Pathway Choices in CRISPR-Cas9-Mediated Genome Editing

“…Loss of the RAD51 paralogs BCDX2 or CX3 significantly reduces RAD51 foci formation in cells, indicating that both are required to promote RAD51 filament formation or stability [146]. Loss of SWS1 or SWSAP1, both components of the human Shu complex, also reduces RAD51 foci formation, although HR efficiency in these cells is comparable to wild-type [147]. These data all suggest roles for BCDX2, CX3, and the Shu complex during RAD51 filament formation, but underlying mechanisms remain unclear.…”

FIRRM cooperates with FIGNL1 to promote RAD51 disassembly during DNA repair

“…Aside from a better understanding of ICL repair pathway choice, several reports have highlighted the complexity of the final stages of ICL repair, such as the resolution of RAD51-mediated strand invasion. For instance, the AAA + adenosine triphosphatase (ATPase) FIGNL1 has been shown to bind to RAD51 and promote its unloading from DNA (9), a function that is limited by the SWSAP1-SWS1-SPIDR complex (9,10). More recently, the helicase HELQ (11) and the HROB-MCM8-MCM9 (12) complex have been shown to contribute to parallel pathways that promote DNA repair synthesis following D-loop formation.…”

RAD51 paralog function in replicative DNA damage and tolerance

“…1; [7,17,18]). Recent investigation of the human Shu complex revealed additional associated factors to include SPIDR and PDS5B [19,20]. SPIDR is a scaffolding protein known to bind RAD51 and the ATPase, FIGNL1 [21][22][23].…”