Rdh54/Tid1 inhibits Rad51-Rad54-mediated D-loop formation and limits D-loop length

Shah, Shanaya Shital; Hartono, Stella R.; Som, Vanessa; Wright, William D.; Chédin, Frédéric; Heyer, Wolf Dietrich

doi:10.7554/elife.59112

Cited by 19 publications

(21 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2 A and B ), although product formation was diminished in the presence of both Rad54 and Rdh54 compared to reactions with just Rad54. This observation was consistent with previous studies showing that the presence of Rdh54 reduces D-loop product formation in reactions with Rad54 and Rad51, albeit the molecular basis for this finding remains unknown ( 70 ). An additional contributing factor to the reduced product formation was that the concentration of Rad54 + Rdh54 was fixed at 90 nM total (i.e., 45 nM Rad54 + 45 nM Rdh54), whereas reactions with just Rad54 contained 90 nM Rad54.…”

Section: Resultssupporting

confidence: 93%

Rad54 and Rdh54 prevent Srs2-mediated disruption of Rad51 presynaptic filaments

Meir

Crickard

Kwon

et al. 2022

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

View full text Add to dashboard Cite

Srs2 is a superfamily 1 (SF1) helicase that participates in several pathways necessary for the repair of damaged DNA. Srs2 regulates formation of early homologous recombination (HR) intermediates by actively removing the recombinase Rad51 from single-stranded DNA (ssDNA). It is not known whether and how Srs2 itself is down-regulated to allow for timely HR progression. Rad54 and Rdh54 are two closely related superfamily 2 (SF2) motor proteins that promote the formation of Rad51-dependent recombination intermediates. Rad54 and Rdh54 bind tightly to Rad51-ssDNA and act downstream of Srs2, suggesting that they may affect the ability of Srs2 to dismantle Rad51 filaments. Here, we used DNA curtains to determine whether Rad54 and Rdh54 alter the ability of Srs2 to disrupt Rad51 filaments. We show that Rad54 and Rdh54 act synergistically to greatly restrict the antirecombinase activity of Srs2. Our findings suggest that Srs2 may be accorded only a limited time window to act and that Rad54 and Rdh54 fulfill a role of prorecombinogenic licensing factors.

show abstract

Section: Resultssupporting

confidence: 93%

Rad54 and Rdh54 prevent Srs2-mediated disruption of Rad51 presynaptic filaments

Meir

Crickard

Kwon

et al. 2022

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

View full text Add to dashboard Cite

show abstract

“…Specific examples of this include the inhibition of Sgs1 translocation by Rad52 [52], the inhibition of Srs2 translocation by Rad54 and Rdh54 [42], and the inhibition of both Srs2 and Sgs1 by the meiosis specific recombinase Dmc1 [53]. Our data indicates that Rdh54 acts as a barrier to Rad54 at newly formed D-loop intermediates, consistent with previous studies that showed Rdh54 limited D-loop size [54]. In contrast to previous reports, we observed that Rdh54 only inhibited the Rad54 mediated D-loop turnover phase of the reaction and did not inhibit D-loop formation.…”

Section: Rdh54 Blocks Rad54 Activity At Newly Formed D-loop Intermedi...supporting

confidence: 89%

Rdh54 stabilizes Rad51 at displacement loop intermediates to regulate genetic exchange between chromosomes

Keymakh

Dau

Ferlez

et al. 2022

Preprint

View full text Add to dashboard Cite

Homologous recombination (HR) is a double-strand break DNA repair pathway that preserves chromosome structure. To repair the damaged recipient DNA, HR requires an intact donor DNA sequence located elsewhere in the genome. After the double-strand break is repaired, DNA sequence information can be transferred between donor and recipient DNA molecules through different mechanisms, including DNA crossovers that form between homologous chromosomes. Regulating this transfer of information is an important step in effectively completing HR and maintaining genome integrity. For example, mitotic exchange of information between homologous chromosomes can result in loss-of-heterozygosity (LOH) in diploid organisms, and in higher eukaryotes, the development of cancer. The DNA motor protein Rdh54 is a highly conserved DNA translocase that functions during HR but has a limited role in repairing DNA. Instead, several existing phenotypes in rdh54∆ strains suggest that Rdh54 may regulate the flow of information between donor and recipient DNA molecules post DNA repair. In our current study, we used a combination of biochemical and genetic techniques to dissect the role of Rdh54 on the exchange of genetic information after DNA repair. Our data indicates that RDH54 regulates DNA sequence exchange between chromosomes by limiting the disruption of Rad51 at an early HR intermediate called the displacement loop (D-loop). Rdh54 also protects Rad51 filaments, acting in opposition to Rad51 removal by the DNA motor protein Rad54. Furthermore, we find that expression of a catalytically inactivate allele of Rdh54, rdh54K318R, displays a different distribution of information exchange outcomes than rdh54∆ cells. From these results, we propose a model for how Rdh54 may effectively regulate information transfer during homologous recombination.

show abstract

“…Previous reports have suggested that Rdh54 competes with Rad54 for D-loop formation. However, these findings were inconclusive in determining whether competition occurred pre-or post-DNA synapsis [60]. Based on our genetic results we hypothesized that post-synaptic inhibition of Rad54 by Rdh54 may support the phenotypes observed in our genetic assay.…”

Section: Rdh54 Regulates D-loop Turnovermentioning

confidence: 81%

Rdh54 stabilizes Rad51 at displacement loop intermediates to regulate genetic exchange between chromosomes

Keymakh

Dau

et al. 2022

PLoS Genet

View full text Add to dashboard Cite

Homologous recombination (HR) is a double-strand break DNA repair pathway that preserves chromosome structure. To repair damaged DNA, HR uses an intact donor DNA sequence located elsewhere in the genome. After the double-strand break is repaired, DNA sequence information can be transferred between donor and recipient DNA molecules through different mechanisms, including DNA crossovers that form between homologous chromosomes. Regulation of DNA sequence transfer is an important step in effectively completing HR and maintaining genome integrity. For example, mitotic exchange of information between homologous chromosomes can result in loss-of-heterozygosity (LOH), and in higher eukaryotes, the development of cancer. The DNA motor protein Rdh54 is a highly conserved DNA translocase that functions during HR. Several existing phenotypes in rdh54Δ strains suggest that Rdh54 may regulate effective exchange of DNA during HR. In our current study, we used a combination of biochemical and genetic techniques to dissect the role of Rdh54 on the exchange of genetic information during DNA repair. Our data indicate that RDH54 regulates DNA strand exchange by stabilizing Rad51 at an early HR intermediate called the displacement loop (D-loop). Rdh54 acts in opposition to Rad51 removal by the DNA motor protein Rad54. Furthermore, we find that expression of a catalytically inactivate allele of Rdh54, rdh54K318R, favors non-crossover outcomes. From these results, we propose a model for how Rdh54 may kinetically regulate strand exchange during homologous recombination.

show abstract

Rdh54/Tid1 inhibits Rad51-Rad54-mediated D-loop formation and limits D-loop length

Cited by 19 publications

References 67 publications

Rad54 and Rdh54 prevent Srs2-mediated disruption of Rad51 presynaptic filaments

Rad54 and Rdh54 prevent Srs2-mediated disruption of Rad51 presynaptic filaments

Rdh54 stabilizes Rad51 at displacement loop intermediates to regulate genetic exchange between chromosomes

Rdh54 stabilizes Rad51 at displacement loop intermediates to regulate genetic exchange between chromosomes

Contact Info

Product

Resources

About