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Pol β associated complex and base excision repair factors in mouse fibroblasts
During mammalian base excision repair (BER) of lesion-containing DNA, it is proposed that toxic strand-break intermediates generated throughout the pathway are sequestered and passed from one step to the next until repair is complete. This stepwise process is termed substrate channeling. A working m...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3526277/ https://www.ncbi.nlm.nih.gov/pubmed/23042675 http://dx.doi.org/10.1093/nar/gks898 |
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author | Prasad, Rajendra Williams, Jason G. Hou, Esther W. Wilson, Samuel H. |
author_facet | Prasad, Rajendra Williams, Jason G. Hou, Esther W. Wilson, Samuel H. |
author_sort | Prasad, Rajendra |
collection | PubMed |
description | During mammalian base excision repair (BER) of lesion-containing DNA, it is proposed that toxic strand-break intermediates generated throughout the pathway are sequestered and passed from one step to the next until repair is complete. This stepwise process is termed substrate channeling. A working model evaluated here is that a complex of BER factors may facilitate the BER process. FLAG-tagged DNA polymerase (pol) β was expressed in mouse fibroblasts carrying a deletion in the endogenous pol β gene, and the cell extract was subjected to an ‘affinity-capture’ procedure using anti-FLAG antibody. The pol β affinity-capture fraction (ACF) was found to contain several BER factors including polymerase-1, X-ray cross-complementing factor1-DNA ligase III and enzymes involved in processing 3′-blocked ends of BER intermediates, e.g. polynucleotide kinase and tyrosyl-DNA phosphodiesterase 1. In contrast, DNA glycosylases, apurinic/aprymidinic endonuclease 1 and flap endonuclease 1 and several other factors involved in BER were not present. Some of the BER factors in the pol β ACF were in a multi-protein complex as observed by sucrose gradient centrifugation. The pol β ACF was capable of substrate channeling for steps in vitro BER and was proficient in in vitro repair of substrates mimicking a 3′-blocked topoisomerase I covalent intermediate or an oxidative stress-induced 3′-blocked intermediate. |
format | Online Article Text |
id | pubmed-3526277 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-35262772013-01-04 Pol β associated complex and base excision repair factors in mouse fibroblasts Prasad, Rajendra Williams, Jason G. Hou, Esther W. Wilson, Samuel H. Nucleic Acids Res Nucleic Acid Enzymes During mammalian base excision repair (BER) of lesion-containing DNA, it is proposed that toxic strand-break intermediates generated throughout the pathway are sequestered and passed from one step to the next until repair is complete. This stepwise process is termed substrate channeling. A working model evaluated here is that a complex of BER factors may facilitate the BER process. FLAG-tagged DNA polymerase (pol) β was expressed in mouse fibroblasts carrying a deletion in the endogenous pol β gene, and the cell extract was subjected to an ‘affinity-capture’ procedure using anti-FLAG antibody. The pol β affinity-capture fraction (ACF) was found to contain several BER factors including polymerase-1, X-ray cross-complementing factor1-DNA ligase III and enzymes involved in processing 3′-blocked ends of BER intermediates, e.g. polynucleotide kinase and tyrosyl-DNA phosphodiesterase 1. In contrast, DNA glycosylases, apurinic/aprymidinic endonuclease 1 and flap endonuclease 1 and several other factors involved in BER were not present. Some of the BER factors in the pol β ACF were in a multi-protein complex as observed by sucrose gradient centrifugation. The pol β ACF was capable of substrate channeling for steps in vitro BER and was proficient in in vitro repair of substrates mimicking a 3′-blocked topoisomerase I covalent intermediate or an oxidative stress-induced 3′-blocked intermediate. Oxford University Press 2012-12 2012-10-05 /pmc/articles/PMC3526277/ /pubmed/23042675 http://dx.doi.org/10.1093/nar/gks898 Text en Published by Oxford University Press 2012. http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial reuse, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com. |
spellingShingle | Nucleic Acid Enzymes Prasad, Rajendra Williams, Jason G. Hou, Esther W. Wilson, Samuel H. Pol β associated complex and base excision repair factors in mouse fibroblasts |
title | Pol β associated complex and base excision repair factors in mouse fibroblasts |
title_full | Pol β associated complex and base excision repair factors in mouse fibroblasts |
title_fullStr | Pol β associated complex and base excision repair factors in mouse fibroblasts |
title_full_unstemmed | Pol β associated complex and base excision repair factors in mouse fibroblasts |
title_short | Pol β associated complex and base excision repair factors in mouse fibroblasts |
title_sort | pol β associated complex and base excision repair factors in mouse fibroblasts |
topic | Nucleic Acid Enzymes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3526277/ https://www.ncbi.nlm.nih.gov/pubmed/23042675 http://dx.doi.org/10.1093/nar/gks898 |
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