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Dna2 is a structure-specific nuclease, with affinity for 5′-flap intermediates
Dna2 is a nuclease/helicase with proposed roles in DNA replication, double-strand break repair and telomere maintenance. For each role Dna2 is proposed to process DNA substrates with a 5′-flap. To date, however, Dna2 has not revealed a preference for binding or cleavage of flaps over single-stranded...
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817469/ https://www.ncbi.nlm.nih.gov/pubmed/19934252 http://dx.doi.org/10.1093/nar/gkp1055 |
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author | Stewart, Jason A. Campbell, Judith L. Bambara, Robert A. |
author_facet | Stewart, Jason A. Campbell, Judith L. Bambara, Robert A. |
author_sort | Stewart, Jason A. |
collection | PubMed |
description | Dna2 is a nuclease/helicase with proposed roles in DNA replication, double-strand break repair and telomere maintenance. For each role Dna2 is proposed to process DNA substrates with a 5′-flap. To date, however, Dna2 has not revealed a preference for binding or cleavage of flaps over single-stranded DNA. Using DNA binding competition assays we found that Dna2 has substrate structure specificity. The nuclease displayed a strong preference for binding substrates with a 5′-flap or some variations of flap structure. Further analysis revealed that Dna2 recognized and bound both the single-stranded flap and portions of the duplex region immediately downstream of the flap. A model is proposed in which Dna2 first binds to a flap base, and then the flap threads through the protein with periodic cleavage, to a terminal flap length of ∼5 nt. This resembles the mechanism of flap endonuclease 1, consistent with cooperation of these two proteins in flap processing. |
format | Text |
id | pubmed-2817469 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-28174692010-02-08 Dna2 is a structure-specific nuclease, with affinity for 5′-flap intermediates Stewart, Jason A. Campbell, Judith L. Bambara, Robert A. Nucleic Acids Res Nucleic Acid Enzymes Dna2 is a nuclease/helicase with proposed roles in DNA replication, double-strand break repair and telomere maintenance. For each role Dna2 is proposed to process DNA substrates with a 5′-flap. To date, however, Dna2 has not revealed a preference for binding or cleavage of flaps over single-stranded DNA. Using DNA binding competition assays we found that Dna2 has substrate structure specificity. The nuclease displayed a strong preference for binding substrates with a 5′-flap or some variations of flap structure. Further analysis revealed that Dna2 recognized and bound both the single-stranded flap and portions of the duplex region immediately downstream of the flap. A model is proposed in which Dna2 first binds to a flap base, and then the flap threads through the protein with periodic cleavage, to a terminal flap length of ∼5 nt. This resembles the mechanism of flap endonuclease 1, consistent with cooperation of these two proteins in flap processing. Oxford University Press 2010-01 2009-11-24 /pmc/articles/PMC2817469/ /pubmed/19934252 http://dx.doi.org/10.1093/nar/gkp1055 Text en © The Author(s) 2009. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5/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/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Nucleic Acid Enzymes Stewart, Jason A. Campbell, Judith L. Bambara, Robert A. Dna2 is a structure-specific nuclease, with affinity for 5′-flap intermediates |
title | Dna2 is a structure-specific nuclease, with affinity for 5′-flap intermediates |
title_full | Dna2 is a structure-specific nuclease, with affinity for 5′-flap intermediates |
title_fullStr | Dna2 is a structure-specific nuclease, with affinity for 5′-flap intermediates |
title_full_unstemmed | Dna2 is a structure-specific nuclease, with affinity for 5′-flap intermediates |
title_short | Dna2 is a structure-specific nuclease, with affinity for 5′-flap intermediates |
title_sort | dna2 is a structure-specific nuclease, with affinity for 5′-flap intermediates |
topic | Nucleic Acid Enzymes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817469/ https://www.ncbi.nlm.nih.gov/pubmed/19934252 http://dx.doi.org/10.1093/nar/gkp1055 |
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