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In cellulo phosphorylation of XRCC4 Ser320 by DNA-PK induced by DNA damage
XRCC4 is a protein associated with DNA Ligase IV, which is thought to join two DNA ends at the final step of DNA double-strand break repair through non-homologous end joining. In response to treatment with ionizing radiation or DNA damaging agents, XRCC4 undergoes DNA-PK-dependent phosphorylation. F...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Oxford University Press
2016
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4795952/ https://www.ncbi.nlm.nih.gov/pubmed/26666690 http://dx.doi.org/10.1093/jrr/rrv086 |
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author | Sharma, Mukesh Kumar Imamichi, Shoji Fukuchi, Mikoto Samarth, Ravindra Mahadeo Tomita, Masanori Matsumoto, Yoshihisa |
author_facet | Sharma, Mukesh Kumar Imamichi, Shoji Fukuchi, Mikoto Samarth, Ravindra Mahadeo Tomita, Masanori Matsumoto, Yoshihisa |
author_sort | Sharma, Mukesh Kumar |
collection | PubMed |
description | XRCC4 is a protein associated with DNA Ligase IV, which is thought to join two DNA ends at the final step of DNA double-strand break repair through non-homologous end joining. In response to treatment with ionizing radiation or DNA damaging agents, XRCC4 undergoes DNA-PK-dependent phosphorylation. Furthermore, Ser260 and Ser320 (or Ser318 in alternatively spliced form) of XRCC4 were identified as the major phosphorylation sites by purified DNA-PK in vitro through mass spectrometry. However, it has not been clear whether these sites are phosphorylated in vivo in response to DNA damage. In the present study, we generated an antibody that reacts with XRCC4 phosphorylated at Ser320 and examined in cellulo phosphorylation status of XRCC4 Ser320. The phosphorylation of XRCC4 Ser320 was induced by γ-ray irradiation and treatment with Zeocin. The phosphorylation of XRCC4 Ser320 was detected even after 1 Gy irradiation and increased in a manner dependent on radiation dose. The phosphorylation was observed immediately after irradiation and remained mostly unchanged for up to 4 h. The phosphorylation was inhibited by DNA-PK inhibitor NU7441 and was undetectable in DNA-PKcs-deficient cells, indicating that the phosphorylation was mainly mediated by DNA-PK. These results suggested potential usefulness of the phosphorylation status of XRCC4 Ser320 as an indicator of DNA-PK functionality in living cells. |
format | Online Article Text |
id | pubmed-4795952 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-47959522016-03-21 In cellulo phosphorylation of XRCC4 Ser320 by DNA-PK induced by DNA damage Sharma, Mukesh Kumar Imamichi, Shoji Fukuchi, Mikoto Samarth, Ravindra Mahadeo Tomita, Masanori Matsumoto, Yoshihisa J Radiat Res Radiation Biology XRCC4 is a protein associated with DNA Ligase IV, which is thought to join two DNA ends at the final step of DNA double-strand break repair through non-homologous end joining. In response to treatment with ionizing radiation or DNA damaging agents, XRCC4 undergoes DNA-PK-dependent phosphorylation. Furthermore, Ser260 and Ser320 (or Ser318 in alternatively spliced form) of XRCC4 were identified as the major phosphorylation sites by purified DNA-PK in vitro through mass spectrometry. However, it has not been clear whether these sites are phosphorylated in vivo in response to DNA damage. In the present study, we generated an antibody that reacts with XRCC4 phosphorylated at Ser320 and examined in cellulo phosphorylation status of XRCC4 Ser320. The phosphorylation of XRCC4 Ser320 was induced by γ-ray irradiation and treatment with Zeocin. The phosphorylation of XRCC4 Ser320 was detected even after 1 Gy irradiation and increased in a manner dependent on radiation dose. The phosphorylation was observed immediately after irradiation and remained mostly unchanged for up to 4 h. The phosphorylation was inhibited by DNA-PK inhibitor NU7441 and was undetectable in DNA-PKcs-deficient cells, indicating that the phosphorylation was mainly mediated by DNA-PK. These results suggested potential usefulness of the phosphorylation status of XRCC4 Ser320 as an indicator of DNA-PK functionality in living cells. Oxford University Press 2016-03 2015-12-13 /pmc/articles/PMC4795952/ /pubmed/26666690 http://dx.doi.org/10.1093/jrr/rrv086 Text en © The Author 2015. Published by Oxford University Press on behalf of The Japan Radiation Research Society and Japanese Society for Radiation Oncology. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Radiation Biology Sharma, Mukesh Kumar Imamichi, Shoji Fukuchi, Mikoto Samarth, Ravindra Mahadeo Tomita, Masanori Matsumoto, Yoshihisa In cellulo phosphorylation of XRCC4 Ser320 by DNA-PK induced by DNA damage |
title | In cellulo phosphorylation of XRCC4 Ser320 by DNA-PK induced by DNA damage |
title_full | In cellulo phosphorylation of XRCC4 Ser320 by DNA-PK induced by DNA damage |
title_fullStr | In cellulo phosphorylation of XRCC4 Ser320 by DNA-PK induced by DNA damage |
title_full_unstemmed | In cellulo phosphorylation of XRCC4 Ser320 by DNA-PK induced by DNA damage |
title_short | In cellulo phosphorylation of XRCC4 Ser320 by DNA-PK induced by DNA damage |
title_sort | in cellulo phosphorylation of xrcc4 ser320 by dna-pk induced by dna damage |
topic | Radiation Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4795952/ https://www.ncbi.nlm.nih.gov/pubmed/26666690 http://dx.doi.org/10.1093/jrr/rrv086 |
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