Saltar al contenido
Merck

MutSβ Stimulates Holliday Junction Resolution by the SMX Complex.

Cell reports (2020-10-22)
Sarah J Young, Marie Sebald, Rajvee Shah Punatar, Meghan Larin, Laura Masino, Monica C Rodrigo-Brenni, Chih-Chao Liang, Stephen C West
RESUMEN

MutSα and MutSβ play important roles in DNA mismatch repair and are linked to inheritable cancers and degenerative disorders. Here, we show that MSH2 and MSH3, the two components of MutSβ, bind SLX4 protein, a scaffold for the assembly of the SLX1-SLX4-MUS81-EME1-XPF-ERCC1 (SMX) trinuclease complex. SMX promotes the resolution of Holliday junctions (HJs), which are intermediates in homologous recombinational repair. We find that MutSβ binds HJs and stimulates their resolution by SLX1-SLX4 or SMX in reactions dependent upon direct interactions between MutSβ and SLX4. In contrast, MutSα does not stimulate HJ resolution. MSH3-depleted cells exhibit reduced sister chromatid exchanges and elevated levels of homologous recombination ultrafine bridges (HR-UFBs) at mitosis, consistent with defects in the processing of recombination intermediates. These results demonstrate a role for MutSβ in addition to its established role in the pathogenic expansion of CAG/CTG trinucleotide repeats, which is causative of myotonic dystrophy and Huntington's disease.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
Alcohol etílico puro, 200 proof, for molecular biology
Sigma-Aldrich
Suero fetal bovino, USA origin, sterile-filtered, suitable for cell culture, suitable for hybridoma
Sigma-Aldrich
Ampicilina sodium salt, powder or crystals, BioReagent, suitable for cell culture
Sigma-Aldrich
Proteinasa K from Tritirachium album, buffered aqueous glycerol solution, for molecular biology, ≥800 units/mL
Sigma-Aldrich
DPX Mountant for histology, slide mounting medium
Sigma-Aldrich
2-Propanol, BioReagent, for molecular biology, ≥99.5%
Sigma-Aldrich
Gentamicina sulfate salt, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Imidazol, for molecular biology, ≥99% (titration)
Sigma-Aldrich
Timidina, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Nocodazole, ≥99% (TLC), powder
Roche
Anti-GFP, from mouse IgG1κ (clones 7.1 and 13.1)
Sigma-Aldrich
Tetracycline, 98.0-102.0% (HPLC)
Sigma-Aldrich
Giemsa stain, certified by the Biological Stain Commission
Roche
Colcemid, solution, suitable for blocking, sterile; 0.2 μm filtered
Sigma-Aldrich
bisBenzimide H 33342 trihydrochloride, ≥98% (HPLC and TLC)
Sigma-Aldrich
p-Xylene-bis(N-pyridinium bromide), ≥95% (TLC)
Sigma-Aldrich
MISSION® esiRNA, targeting human GEN1
Sigma-Aldrich
MISSION® esiRNA, targeting human MSH3
Sigma-Aldrich
MISSION® esiRNA, targeting human MSH6