Accéder au contenu
Merck

Down-regulation of Rad51 activity during meiosis in yeast prevents competition with Dmc1 for repair of double-strand breaks.

PLoS genetics (2014-01-28)
Yan Liu, William A Gaines, Tracy Callender, Valeria Busygina, Ashwini Oke, Patrick Sung, Jennifer C Fung, Nancy M Hollingsworth
RÉSUMÉ

Interhomolog recombination plays a critical role in promoting proper meiotic chromosome segregation but a mechanistic understanding of this process is far from complete. In vegetative cells, Rad51 is a highly conserved recombinase that exhibits a preference for repairing double strand breaks (DSBs) using sister chromatids, in contrast to the conserved, meiosis-specific recombinase, Dmc1, which preferentially repairs programmed DSBs using homologs. Despite the different preferences for repair templates, both Rad51 and Dmc1 are required for interhomolog recombination during meiosis. This paradox has recently been explained by the finding that Rad51 protein, but not its strand exchange activity, promotes Dmc1 function in budding yeast. Rad51 activity is inhibited in dmc1Δ mutants, where the failure to repair meiotic DSBs triggers the meiotic recombination checkpoint, resulting in prophase arrest. The question remains whether inhibition of Rad51 activity is important during wild-type meiosis, or whether inactivation of Rad51 occurs only as a result of the absence of DMC1 or checkpoint activation. This work shows that strains in which mechanisms that down-regulate Rad51 activity are removed exhibit reduced numbers of interhomolog crossovers and noncrossovers. A hypomorphic mutant, dmc1-T159A, makes less stable presynaptic filaments but is still able to mediate strand exchange and interact with accessory factors. Combining dmc1-T159A with up-regulated Rad51 activity reduces interhomolog recombination and spore viability, while increasing intersister joint molecule formation. These results support the idea that down-regulation of Rad51 activity is important during meiosis to prevent Rad51 from competing with Dmc1 for repair of meiotic DSBs.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Phosphate de potassium dibasic, ACS reagent, ≥98%
Sigma-Aldrich
2-mercaptoéthanol, for molecular biology, suitable for electrophoresis, suitable for cell culture, BioReagent, 99% (GC/titration)
Sigma-Aldrich
2-mercaptoéthanol, ≥99.0%
Sigma-Aldrich
Chlorure de magnésium, anhydrous, ≥98%
Sigma-Aldrich
Chlorure de magnésium solution, for molecular biology, 1.00 M±0.01 M
Sigma-Aldrich
Phosphate de potassium dibasic, puriss. p.a., ACS reagent, anhydrous, ≥99.0% (T)
Sigma-Aldrich
2-mercaptoéthanol, BioUltra, for molecular biology, ≥99.0% (GC)
Sigma-Aldrich
Chlorure de magnésium, powder, <200 μm
Sigma-Aldrich
Phosphate de potassium dibasic, anhydrous, for luminescence, for molecular biology, BioUltra, ≥99.0% (T)
Sigma-Aldrich
Chlorure de magnésium solution, BioUltra, for molecular biology, 2 M in H2O
Sigma-Aldrich
Chlorure de magnésium, BioReagent, suitable for insect cell culture, ≥97.0%
Sigma-Aldrich
Potassium phosphate dibasic solution, 1.0 M
Sigma-Aldrich
Chlorure de magnésium solution, PCR Reagent, 25 mM MgCI2 solution for PCR
Sigma-Aldrich
Chlorure de magnésium, AnhydroBeads, −10 mesh, 99.9% trace metals basis
Sigma-Aldrich
Phosphate de potassium dibasic, reagent grade, ≥98.0%
Sigma-Aldrich
Phosphate de potassium dibasic, 99.95% trace metals basis
Sigma-Aldrich
Chlorure de magnésium solution, BioUltra, for molecular biology, ~1 M in H2O
Sigma-Aldrich
Chlorure de magnésium, AnhydroBeads, −10 mesh, 99.99% trace metals basis
Sigma-Aldrich
Chlorure de magnésium solution, 0.1 M
Sigma-Aldrich
Potassium phosphate dibasic, meets USP testing specifications
Supelco
2-mercaptoéthanol, for HPLC derivatization, LiChropur, ≥99.0% (GC)
Sigma-Aldrich
Chlorure de magnésium solution, BioUltra, for molecular biology, ~0.025 M in H2O