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Coordinate 5′ and 3′ endonucleolytic trimming of terminally blocked blunt DNA double-strand break ends by Artemis nuclease and DNA-dependent protein kinase
Previous work showed that, in the presence of DNA-dependent protein kinase (DNA-PK), Artemis slowly trims 3′-phosphoglycolate-terminated blunt ends. To examine the trimming reaction in more detail, long internally labeled DNA substrates were treated with Artemis. In the absence of DNA-PK, Artemis ca...
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Oxford University Press
2008
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| Acesso em linha: | https://ncbi.nlm.nih.gov/pmc/articles/PMC2425473/ https://ncbi.nlm.nih.gov/pubmed/18440975 https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1093/nar/gkn205 |
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pubmed-24254732008-06-12 Coordinate 5′ and 3′ endonucleolytic trimming of terminally blocked blunt DNA double-strand break ends by Artemis nuclease and DNA-dependent protein kinase Yannone, Steven M. Khan, Imran S. Zhou, Rui-Zhe Zhou, Tong Valerie, Kristoffer Povirk, Lawrence F. Nucleic Acids Res Nucleic Acid Enzymes Previous work showed that, in the presence of DNA-dependent protein kinase (DNA-PK), Artemis slowly trims 3′-phosphoglycolate-terminated blunt ends. To examine the trimming reaction in more detail, long internally labeled DNA substrates were treated with Artemis. In the absence of DNA-PK, Artemis catalyzed extensive 5′→3′ exonucleolytic resection of double-stranded DNA. This resection required a 5′-phosphate, but did not require ATP, and was accompanied by endonucleolytic cleavage of the resulting 3′ overhang. In the presence of DNA-PK, Artemis-mediated trimming was more limited, was ATP-dependent and did not require a 5′-phosphate. For a blunt end with either a 3′-phosphoglycolate or 3′-hydroxyl terminus, endonucleolytic trimming of 2–4 nucleotides from the 3′-terminal strand was accompanied by trimming of 6 nt from the 5′-terminal strand. The results suggest that autophosphorylated DNA-PK suppresses the exonuclease activity of Artemis toward blunt-ended DNA, and promotes slow and limited endonucleolytic trimming of the 5′-terminal strand, resulting in short 3′ overhangs that are trimmed endonucleolytically. Thus, Artemis and DNA-PK can convert terminally blocked DNA ends of diverse geometry and chemical structure to a form suitable for polymerase-mediated patching and ligation, with minimal loss of terminal sequence. Such processing could account for the very small deletions often found at DNA double-strand break repair sites. Oxford University Press 2008-06 2008-04-25 /pmc/articles/PMC2425473/ /pubmed/18440975 http://dx.doi.org/10.1093/nar/gkn205 Text en © 2008 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
| institution |
US National Library of Medicine |
| collection |
PubMed Central |
| language |
English |
| format |
Article |
| topic |
Nucleic Acid Enzymes |
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Nucleic Acid Enzymes Yannone, Steven M. Khan, Imran S. Zhou, Rui-Zhe Zhou, Tong Valerie, Kristoffer Povirk, Lawrence F. Coordinate 5′ and 3′ endonucleolytic trimming of terminally blocked blunt DNA double-strand break ends by Artemis nuclease and DNA-dependent protein kinase |
| description |
Previous work showed that, in the presence of DNA-dependent protein kinase (DNA-PK), Artemis slowly trims 3′-phosphoglycolate-terminated blunt ends. To examine the trimming reaction in more detail, long internally labeled DNA substrates were treated with Artemis. In the absence of DNA-PK, Artemis catalyzed extensive 5′→3′ exonucleolytic resection of double-stranded DNA. This resection required a 5′-phosphate, but did not require ATP, and was accompanied by endonucleolytic cleavage of the resulting 3′ overhang. In the presence of DNA-PK, Artemis-mediated trimming was more limited, was ATP-dependent and did not require a 5′-phosphate. For a blunt end with either a 3′-phosphoglycolate or 3′-hydroxyl terminus, endonucleolytic trimming of 2–4 nucleotides from the 3′-terminal strand was accompanied by trimming of 6 nt from the 5′-terminal strand. The results suggest that autophosphorylated DNA-PK suppresses the exonuclease activity of Artemis toward blunt-ended DNA, and promotes slow and limited endonucleolytic trimming of the 5′-terminal strand, resulting in short 3′ overhangs that are trimmed endonucleolytically. Thus, Artemis and DNA-PK can convert terminally blocked DNA ends of diverse geometry and chemical structure to a form suitable for polymerase-mediated patching and ligation, with minimal loss of terminal sequence. Such processing could account for the very small deletions often found at DNA double-strand break repair sites. |
| author |
Yannone, Steven M. Khan, Imran S. Zhou, Rui-Zhe Zhou, Tong Valerie, Kristoffer Povirk, Lawrence F. |
| author_facet |
Yannone, Steven M. Khan, Imran S. Zhou, Rui-Zhe Zhou, Tong Valerie, Kristoffer Povirk, Lawrence F. |
| author_sort |
Yannone, Steven M. |
| title |
Coordinate 5′ and 3′ endonucleolytic trimming of terminally blocked blunt DNA double-strand break ends by Artemis nuclease and DNA-dependent protein kinase |
| title_short |
Coordinate 5′ and 3′ endonucleolytic trimming of terminally blocked blunt DNA double-strand break ends by Artemis nuclease and DNA-dependent protein kinase |
| title_full |
Coordinate 5′ and 3′ endonucleolytic trimming of terminally blocked blunt DNA double-strand break ends by Artemis nuclease and DNA-dependent protein kinase |
| title_fullStr |
Coordinate 5′ and 3′ endonucleolytic trimming of terminally blocked blunt DNA double-strand break ends by Artemis nuclease and DNA-dependent protein kinase |
| title_full_unstemmed |
Coordinate 5′ and 3′ endonucleolytic trimming of terminally blocked blunt DNA double-strand break ends by Artemis nuclease and DNA-dependent protein kinase |
| title_sort |
coordinate 5′ and 3′ endonucleolytic trimming of terminally blocked blunt dna double-strand break ends by artemis nuclease and dna-dependent protein kinase |
| publisher |
Oxford University Press |
| publisher_facet |
Oxford University Press |
| publishDate |
2008 |
| url |
https://ncbi.nlm.nih.gov/pmc/articles/PMC2425473/ https://ncbi.nlm.nih.gov/pubmed/18440975 https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1093/nar/gkn205 |
| _version_ |
1760720157901062144 |