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Chemical mapping of cytosines enzymatically flipped out of the DNA helix
Haloacetaldehydes can be employed for probing unpaired DNA structures involving cytosine and adenine residues. Using an enzyme that was structurally proven to flip its target cytosine out of the DNA helix, the HhaI DNA methyltransferase (M.HhaI), we demonstrate the suitability of the chloroacetaldeh...
Autores principales: | , , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2425465/ https://www.ncbi.nlm.nih.gov/pubmed/18450817 http://dx.doi.org/10.1093/nar/gkn200 |
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author | Daujotytė, Dalia Liutkevičiūtė, Zita Tamulaitis, Gintautas Klimašauskas, Saulius |
author_facet | Daujotytė, Dalia Liutkevičiūtė, Zita Tamulaitis, Gintautas Klimašauskas, Saulius |
author_sort | Daujotytė, Dalia |
collection | PubMed |
description | Haloacetaldehydes can be employed for probing unpaired DNA structures involving cytosine and adenine residues. Using an enzyme that was structurally proven to flip its target cytosine out of the DNA helix, the HhaI DNA methyltransferase (M.HhaI), we demonstrate the suitability of the chloroacetaldehyde modification for mapping extrahelical (flipped-out) cytosine bases in protein–DNA complexes. The generality of this method was verified with two other DNA cytosine-5 methyltransferases, M.AluI and M.SssI, as well as with two restriction endonucleases, R.Ecl18kI and R.PspGI, which represent a novel class of base-flipping enzymes. Our results thus offer a simple and convenient laboratory tool for detection and mapping of flipped-out cytosines in protein–DNA complexes. |
format | Text |
id | pubmed-2425465 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-24254652008-06-12 Chemical mapping of cytosines enzymatically flipped out of the DNA helix Daujotytė, Dalia Liutkevičiūtė, Zita Tamulaitis, Gintautas Klimašauskas, Saulius Nucleic Acids Res Methods Online Haloacetaldehydes can be employed for probing unpaired DNA structures involving cytosine and adenine residues. Using an enzyme that was structurally proven to flip its target cytosine out of the DNA helix, the HhaI DNA methyltransferase (M.HhaI), we demonstrate the suitability of the chloroacetaldehyde modification for mapping extrahelical (flipped-out) cytosine bases in protein–DNA complexes. The generality of this method was verified with two other DNA cytosine-5 methyltransferases, M.AluI and M.SssI, as well as with two restriction endonucleases, R.Ecl18kI and R.PspGI, which represent a novel class of base-flipping enzymes. Our results thus offer a simple and convenient laboratory tool for detection and mapping of flipped-out cytosines in protein–DNA complexes. Oxford University Press 2008-06 2008-05-01 /pmc/articles/PMC2425465/ /pubmed/18450817 http://dx.doi.org/10.1093/nar/gkn200 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. |
spellingShingle | Methods Online Daujotytė, Dalia Liutkevičiūtė, Zita Tamulaitis, Gintautas Klimašauskas, Saulius Chemical mapping of cytosines enzymatically flipped out of the DNA helix |
title | Chemical mapping of cytosines enzymatically flipped out of the DNA helix |
title_full | Chemical mapping of cytosines enzymatically flipped out of the DNA helix |
title_fullStr | Chemical mapping of cytosines enzymatically flipped out of the DNA helix |
title_full_unstemmed | Chemical mapping of cytosines enzymatically flipped out of the DNA helix |
title_short | Chemical mapping of cytosines enzymatically flipped out of the DNA helix |
title_sort | chemical mapping of cytosines enzymatically flipped out of the dna helix |
topic | Methods Online |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2425465/ https://www.ncbi.nlm.nih.gov/pubmed/18450817 http://dx.doi.org/10.1093/nar/gkn200 |
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