Preventing oxidation of cellular XRCC1 affects PARP-mediated DNA damage responses

Autor: Robert E. London, Padmini S. Kedar, Donna F. Stefanick, Scott A. Gabel, Jason Williams, Rajendra Prasad, Eugene F. DeRose, Julie K. Horton, Samuel H. Wilson, Natalie R. Gassman, Matthew J. Cuneo, Esther W. Hou
Rok vydání: 2013
Předmět:
Models
Molecular
Poly Adenosine Diphosphate Ribose
DNA Repair
DNA repair
DNA polymerase
DNA damage
Poly ADP ribose polymerase
Mutant
Mutation
Missense
Poly (ADP-Ribose) Polymerase-1
Poly(ADP-ribose) Polymerase Inhibitors
Quinolones
Biochemistry
Article
Inhibitory Concentration 50
Mice
chemistry.chemical_compound
Animals
Transition Temperature
Antineoplastic Agents
Alkylating
Nuclear Magnetic Resonance
Biomolecular
Molecular Biology
Cells
Cultured
DNA Polymerase beta
Mice
Knockout
biology
Cell Cycle
Hydrogen Peroxide
Cell Biology
Base excision repair
Methyl Methanesulfonate
Oxidants
Molecular biology
Protein Structure
Tertiary
Methyl methanesulfonate
DNA-Binding Proteins
Naphthalimides
1-Naphthylamine
X-ray Repair Cross Complementing Protein 1
chemistry
PARP inhibitor
biology.protein
Poly(ADP-ribose) Polymerases
Oxidation-Reduction
DNA Damage
Protein Binding
Zdroj: DNA Repair. 12:774-785
ISSN: 1568-7864
DOI: 10.1016/j.dnarep.2013.06.004
Popis: Poly(ADP-ribose) polymerase-1 (PARP-1) binds intermediates of base excision repair (BER) and becomes activated for poly(ADP-ribose) (PAR) synthesis. PAR mediates recruitment and functions of the key BER factors XRCC1 and DNA polymerase β (pol β) that in turn regulate PAR. Yet, the molecular mechanism and implications of coordination between XRCC1 and pol β in regulating the level of PAR are poorly understood. A complex of PARP-1, XRCC1 and pol β is found in vivo, and it is known that pol β and XRCC1 interact through a redox-sensitive binding interface in the N-terminal domain of XRCC1. We confirmed here that both oxidized and reduced forms of XRCC1 are present in mouse fibroblasts. To further understand the importance of the C12-C20 oxidized form of XRCC1 and the interaction with pol β, we characterized cell lines representing stable transfectants in Xrcc1(-/-) mouse fibroblasts of wild-type XRCC1 and two mutants of XRCC1, a novel reduced form with the C12-C20 disulfide bond blocked (C12A) and a reference mutant that is unable to bind pol β (V88R). XRCC1-deficient mouse fibroblasts are extremely hypersensitive to methyl methanesulfonate (MMS), and transfected wild-type and C12A mutant XRCC1 proteins similarly reversed MMS hypersensitivity. However, after MMS exposure the cellular PAR level was found to increase to a much greater extent in cells expressing the C12A mutant than in cells expressing wild-type XRCC1. PARP inhibition resulted in very strong MMS sensitization in cells expressing wild-type XRCC1, but this sensitization was much less in cells expressing the C12A mutant. The results suggest a role for the oxidized form of XRCC1 in the interaction with pol β in (1) controlling the PAR level after MMS exposure and (2) enabling the extreme cytotoxicity of PARP inhibition during the MMS DNA damage response.
Databáze: OpenAIRE
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