Skip to Content
Merck
  • MCM8IP activates the MCM8-9 helicase to promote DNA synthesis and homologous recombination upon DNA damage.

MCM8IP activates the MCM8-9 helicase to promote DNA synthesis and homologous recombination upon DNA damage.

Nature communications (2020-06-13)
Jen-Wei Huang, Ananya Acharya, Angelo Taglialatela, Tarun S Nambiar, Raquel Cuella-Martin, Giuseppe Leuzzi, Samuel B Hayward, Sarah A Joseph, Gregory J Brunette, Roopesh Anand, Rajesh K Soni, Nathan L Clark, Kara A Bernstein, Petr Cejka, Alberto Ciccia
ABSTRACT

Homologous recombination (HR) mediates the error-free repair of DNA double-strand breaks to maintain genomic stability. Here we characterize C17orf53/MCM8IP, an OB-fold containing protein that binds ssDNA, as a DNA repair factor involved in HR. MCM8IP-deficient cells exhibit HR defects, especially in long-tract gene conversion, occurring downstream of RAD51 loading, consistent with a role for MCM8IP in HR-dependent DNA synthesis. Moreover, loss of MCM8IP confers cellular sensitivity to crosslinking agents and PARP inhibition. Importantly, we report that MCM8IP directly associates with MCM8-9, a helicase complex mutated in primary ovarian insufficiency, and RPA1. We additionally show that the interactions of MCM8IP with MCM8-9 and RPA facilitate HR and promote replication fork progression and cellular viability in response to treatment with crosslinking agents. Mechanistically, MCM8IP stimulates the helicase activity of MCM8-9. Collectively, our work identifies MCM8IP as a key regulator of MCM8-9-dependent DNA synthesis during DNA recombination and replication.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-HA antibody, Mouse monoclonal, clone HA-7, purified from hybridoma cell culture
Sigma-Aldrich
Monoclonal Anti-Vinculin antibody produced in mouse, clone hVIN-1, ascites fluid
Sigma-Aldrich
3X FLAG® Peptide, lyophilized powder
Sigma-Aldrich
Geranyl pyrophosphate ammonium salt, 1 mg/mL in methanol (:aqueous 10 mM NH4OH (7:3)), ≥95% (TLC)
Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
Sigma-Aldrich
Maltose solution, for molecular biology, BioReagent, ~20% in H2O
Sigma-Aldrich
Anti-C17orf53 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution, Ab2
Millipore
ANTI-FLAG® M2 Affinity Gel, purified immunoglobulin, buffered aqueous glycerol solution
Millipore
Monoclonal Anti-HA−Agarose antibody produced in mouse, clone HA-7, purified immunoglobulin, PBS suspension