PloS one, 2013-06, Vol.8 (6), p.e66801
2013
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- Autor(en) / Beteiligte
- Titel
- Interaction between DNA Polymerase β and BRCA1
- Ist Teil von
- PloS one, 2013-06, Vol.8 (6), p.e66801
- Ort / Verlag
- United States: Public Library of Science
- Erscheinungsjahr
- 2013
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- The breast cancer 1 (BRCA1) protein is a tumor suppressor playing roles in DNA repair and cell cycle regulation. Studies of DNA repair functions of BRCA1 have focused on double-strand break (DSB) repair pathways and have recently included base excision repair (BER). However, the function of BRCA1 in BER is not well defined. Here, we examined a BRCA1 role in BER, first in relation to alkylating agent (MMS) treatment of cells and the BER enzyme DNA polymerase β (pol β). MMS treatment of BRCA1 negative human ovarian and chicken DT40 cells revealed hypersensitivity, and the combined gene deletion of BRCA1 and pol β in DT40 cells was consistent with these factors acting in the same repair pathway, possibly BER. Using cell extracts and purified proteins, BRCA1 and pol β were found to interact in immunoprecipitation assays, yet in vivo and in vitro assays for a BER role of BRCA1 were negative. An alternate approach with the human cells of immunofluorescence imaging and laser-induced DNA damage revealed negligible BRCA1 recruitment during the first 60 s after irradiation, the period typical of recruitment of pol β and other BER factors. Instead, 15 min after irradiation, BRCA1 recruitment was strong and there was γ-H2AX co-localization, consistent with DSBs and repair. The rapid recruitment of pol β was similar in BRCA1 positive and negative cells. However, a fraction of pol β initially recruited remained associated with damage sites much longer in BRCA1 positive than negative cells. Interestingly, pol β expression was required for BRCA1 recruitment, suggesting a partnership between these repair factors in DSB repair.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1932-6203eISSN: 1932-6203DOI: 10.1371/journal.pone.0066801Titel-ID: cdi_plos_journals_1372124281
- Format
- –
- Schlagworte
- Alkylating Agents - toxicity, Alkylation, Animals, Antigens, Base excision repair, Biology, BRCA1 protein, BRCA1 Protein - genetics, BRCA1 Protein - metabolism, Breast cancer, Cell cycle, Cell Line, Tumor, Chickens, Clonal deletion, Deoxyribonucleic acid, DNA, DNA Breaks, Double-Stranded, DNA damage, DNA methylation, DNA polymerase, DNA Polymerase beta - genetics, DNA Polymerase beta - metabolism, DNA repair, DNA Repair - physiology, DNA-directed DNA polymerase, Double-strand break repair, Enzymes, Fluorescent Antibody Technique, Gene deletion, Gene Knockdown Techniques, Histones - metabolism, Humans, Hypersensitivity, Immunofluorescence, Immunoprecipitation, Irradiation, Laboratories, Laser damage, Localization, Medicine, Ovarian cancer, Oxidative stress, Proteins, Radiation damage, Recruitment, Repair, RNA, Small Interfering, Toxicology, Tumor suppressor genes