Premium
Integrin engagement reduces APE1 co‐immunoprecipitation with XRCC1 in hydrogen peroxide‐treated murine lung endothelial cells
Author(s) -
Stevens Rachel L.,
Hoyt Dale G.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.777.2
Subject(s) - microbiology and biotechnology , dna damage , integrin , immunoprecipitation , xrcc1 , integrin alpha m , phosphorylation , chemistry , antibody , biology , immunology , dna , cell , biochemistry , flow cytometry , gene , genotype , single nucleotide polymorphism
Reduction of genotoxin‐induced DNA 3’OH end labeling and histone 2A.X (H2A.X) phosphorylation after integrin engagement requires certain Base Excision Repair (BER) proteins; however, integrin engagement does not reduce the amount of physical DNA breaks as measured by the comet assay. Consequently, we hypothesized that integrin signaling mediates the outcome of specific processing events during the BER response. To address this, murine lung endothelial cells were plated overnight in DMEM with 0.5% FBS and treated with 1 μg anti‐β1 integrin antibody/ml or the corresponding non‐immune IgG for 1 h followed by addition of 2 μg goat anti‐rat secondary antibody/ml for 4 h. Cells were then treated with and without 0.001M H 2 O 2 for 15 min. to induce DNA damage. XRCC1 was successfully immunoprecipitated (IP) after incubating cell lysate with XRCC1 antibody overnight. Subsequent Western blotting for APE1 revealed that the non‐immune IgG‐treated samples damaged with H 2 O 2 exhibited the expected increase in APE1‐XRCC1 association by IP. By contrast, integrin engagement significantly reduced co‐IP of APE1 with XRCC1 after H 2 O 2 ‐induced damage. These results suggest that integrin signaling modulates the response to DNA breakage through altered BER protein interactions, and may account for the previously observed reductions in DNA 3’OH and H2A.X phosphorylation.