Skip to Content
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
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

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