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Kinetics of endogenous mouse FEN1 in base excision repair
The structure specific flap endonuclease 1 (FEN1) plays an essential role in long-patch base excision repair (BER) and in DNA replication. We have generated a fluorescently tagged FEN1 expressing mouse which allows monitoring the localization and kinetics of FEN1 in response to DNA damage in living...
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/PMC3467068/ https://www.ncbi.nlm.nih.gov/pubmed/22810208 http://dx.doi.org/10.1093/nar/gks673 |
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author | Kleppa, Liv Mari, Pierre-Olivier Larsen, Elisabeth Lien, Guro Flor Godon, Camille Theil, Arjan F. Nesse, Gaute J. Wiksen, Hege Vermeulen, Wim Giglia-Mari, Giuseppina Klungland, Arne |
author_facet | Kleppa, Liv Mari, Pierre-Olivier Larsen, Elisabeth Lien, Guro Flor Godon, Camille Theil, Arjan F. Nesse, Gaute J. Wiksen, Hege Vermeulen, Wim Giglia-Mari, Giuseppina Klungland, Arne |
author_sort | Kleppa, Liv |
collection | PubMed |
description | The structure specific flap endonuclease 1 (FEN1) plays an essential role in long-patch base excision repair (BER) and in DNA replication. We have generated a fluorescently tagged FEN1 expressing mouse which allows monitoring the localization and kinetics of FEN1 in response to DNA damage in living cells and tissues. The expression of FEN1, which is tagged at its C-terminal end with enhanced yellow fluorescent protein (FEN1-YFP), is under control of the endogenous Fen1 transcriptional regulatory elements. In line with its role in processing of Okazaki fragments during DNA replication, we found that FEN1-YFP expression is mainly observed in highly proliferating tissue. Moreover, the FEN1-YFP fusion protein allowed us to investigate repair kinetics in cells challenged with local and global DNA damage. In vivo multi-photon fluorescence microscopy demonstrates rapid localization of FEN1 to local laser-induced DNA damage sites in nuclei, providing evidence of a highly mobile protein that accumulates fast at DNA lesion sites with high turnover rate. Inhibition of poly (ADP-ribose) polymerase 1 (PARP1) disrupts FEN1 accumulation at sites of DNA damage, indicating that PARP1 is required for FEN1 recruitment to DNA repair intermediates in BER. |
format | Online Article Text |
id | pubmed-3467068 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-34670682012-10-10 Kinetics of endogenous mouse FEN1 in base excision repair Kleppa, Liv Mari, Pierre-Olivier Larsen, Elisabeth Lien, Guro Flor Godon, Camille Theil, Arjan F. Nesse, Gaute J. Wiksen, Hege Vermeulen, Wim Giglia-Mari, Giuseppina Klungland, Arne Nucleic Acids Res Genome Integrity, Repair and Replication The structure specific flap endonuclease 1 (FEN1) plays an essential role in long-patch base excision repair (BER) and in DNA replication. We have generated a fluorescently tagged FEN1 expressing mouse which allows monitoring the localization and kinetics of FEN1 in response to DNA damage in living cells and tissues. The expression of FEN1, which is tagged at its C-terminal end with enhanced yellow fluorescent protein (FEN1-YFP), is under control of the endogenous Fen1 transcriptional regulatory elements. In line with its role in processing of Okazaki fragments during DNA replication, we found that FEN1-YFP expression is mainly observed in highly proliferating tissue. Moreover, the FEN1-YFP fusion protein allowed us to investigate repair kinetics in cells challenged with local and global DNA damage. In vivo multi-photon fluorescence microscopy demonstrates rapid localization of FEN1 to local laser-induced DNA damage sites in nuclei, providing evidence of a highly mobile protein that accumulates fast at DNA lesion sites with high turnover rate. Inhibition of poly (ADP-ribose) polymerase 1 (PARP1) disrupts FEN1 accumulation at sites of DNA damage, indicating that PARP1 is required for FEN1 recruitment to DNA repair intermediates in BER. Oxford University Press 2012-10 2012-07-18 /pmc/articles/PMC3467068/ /pubmed/22810208 http://dx.doi.org/10.1093/nar/gks673 Text en © The Author(s) 2012. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Genome Integrity, Repair and Replication Kleppa, Liv Mari, Pierre-Olivier Larsen, Elisabeth Lien, Guro Flor Godon, Camille Theil, Arjan F. Nesse, Gaute J. Wiksen, Hege Vermeulen, Wim Giglia-Mari, Giuseppina Klungland, Arne Kinetics of endogenous mouse FEN1 in base excision repair |
title | Kinetics of endogenous mouse FEN1 in base excision repair |
title_full | Kinetics of endogenous mouse FEN1 in base excision repair |
title_fullStr | Kinetics of endogenous mouse FEN1 in base excision repair |
title_full_unstemmed | Kinetics of endogenous mouse FEN1 in base excision repair |
title_short | Kinetics of endogenous mouse FEN1 in base excision repair |
title_sort | kinetics of endogenous mouse fen1 in base excision repair |
topic | Genome Integrity, Repair and Replication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3467068/ https://www.ncbi.nlm.nih.gov/pubmed/22810208 http://dx.doi.org/10.1093/nar/gks673 |
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