Abstract
Microcephalin (MCPH1) is one of the causative genes for the autosomal recessive disorder, primary microcephaly, characterized by dramatic reduction in brain size and mental retardation. MCPH1 also functions in the DNA damage response, participating in cell cycle checkpoint control. However, how MCPH1 is regulated in the DNA damage response still remains unknown. Here we report that the ability of MCPH1 to localize to the sites of DNA double-strand breaks depends on its C-terminal tandem BRCT domains. Although MCPH1 foci formation depends on H2AX phosphorylation after DNA damage, it can occur independently of MDC1. We also show that MCPH1 binds to a phospho-H2AX peptide in vitro with an affinity similar to that of MDC1, and overexpression of wild type, but not C-BRCT mutants of MCPH1, can interfere with the foci formation of MDC1 and 53BP1. Collectively, our data suggest MCPH1 is recruited to double-strand breaks via its interaction with gammaH2AX, which is mediated by MCPH1 C-terminal BRCT domains. These observations support that MCPH1 acts early in DNA damage responsive pathways.
MeSH Terms
Adaptor Proteins, Signal Transducing
Cell Cycle Proteins
Cell Line
Cell Nucleus/metabolism
Cell Separation
Cytoskeletal Proteins
DNA Damage
DNA Repair
DNA-Binding Proteins/metabolism
Flow Cytometry
HeLa Cells
Histones/metabolism
Humans
Kinetics
Models, Biological
Mutation
Nerve Tissue Proteins/metabolism,physiology
Nuclear Proteins/metabolism
Phosphorylation
Trans-Activators/metabolism
Chemicals
Adaptor Proteins, Signal Transducing
Cell Cycle Proteins
Cytoskeletal Proteins
DNA-Binding Proteins
H2AX protein, human
Histones
MCPH1 protein, human
MDC1 protein, human
Nerve Tissue Proteins
Nuclear Proteins
Trans-Activators
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wood Jamie L
Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic College of Medicine, Rochester, Minnesota 55905, USA.
Singh Namit
Mer Georges
Chen Junjie
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