Restriction endonuclease MvaI is a monomer that recognizes its target sequence asymmetrically

Restriction endonuclease MvaI recognizes the sequence CC/WGG (W stands for A or T, ‘/’ designates the cleavage site) and generates products with single nucleotide 5′-overhangs. The enzyme has been noted for its tolerance towards DNA modifications. Here, we report a biochemical characterization and c...

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Autores principales: Kaus-Drobek, Magdalena, Czapinska, Honorata, Sokołowska, Monika, Tamulaitis, Gintautas, Szczepanowski, Roman H., Urbanke, Claus, Siksnys, Virginijus, Bochtler, Matthias
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1874612/
https://www.ncbi.nlm.nih.gov/pubmed/17344322
http://dx.doi.org/10.1093/nar/gkm064
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author Kaus-Drobek, Magdalena
Czapinska, Honorata
Sokołowska, Monika
Tamulaitis, Gintautas
Szczepanowski, Roman H.
Urbanke, Claus
Siksnys, Virginijus
Bochtler, Matthias
author_facet Kaus-Drobek, Magdalena
Czapinska, Honorata
Sokołowska, Monika
Tamulaitis, Gintautas
Szczepanowski, Roman H.
Urbanke, Claus
Siksnys, Virginijus
Bochtler, Matthias
author_sort Kaus-Drobek, Magdalena
collection PubMed
description Restriction endonuclease MvaI recognizes the sequence CC/WGG (W stands for A or T, ‘/’ designates the cleavage site) and generates products with single nucleotide 5′-overhangs. The enzyme has been noted for its tolerance towards DNA modifications. Here, we report a biochemical characterization and crystal structures of MvaI in an apo-form and in a complex with target DNA at 1.5 Å resolution. Our results show that MvaI is a monomer and recognizes its pseudosymmetric target sequence asymmetrically. The enzyme consists of two lobes. The catalytic lobe anchors the active site residues Glu36, Asp50, Glu55 and Lys57 and contacts the bases from the minor grove side. The recognition lobe mediates all major grove interactions with the bases. The enzyme in the crystal is bound to the strand with T at the center of the recognition sequence. The crystal structure with calcium ions and DNA mimics the prereactive state. MvaI shows structural similarities to BcnI, which cleaves the related sequence CC/SGG and to MutH enzyme, which is a component of the DNA repair machinery, and nicks one DNA strand instead of making a double-strand break.
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spelling pubmed-18746122007-05-23 Restriction endonuclease MvaI is a monomer that recognizes its target sequence asymmetrically Kaus-Drobek, Magdalena Czapinska, Honorata Sokołowska, Monika Tamulaitis, Gintautas Szczepanowski, Roman H. Urbanke, Claus Siksnys, Virginijus Bochtler, Matthias Nucleic Acids Res Structural Biology Restriction endonuclease MvaI recognizes the sequence CC/WGG (W stands for A or T, ‘/’ designates the cleavage site) and generates products with single nucleotide 5′-overhangs. The enzyme has been noted for its tolerance towards DNA modifications. Here, we report a biochemical characterization and crystal structures of MvaI in an apo-form and in a complex with target DNA at 1.5 Å resolution. Our results show that MvaI is a monomer and recognizes its pseudosymmetric target sequence asymmetrically. The enzyme consists of two lobes. The catalytic lobe anchors the active site residues Glu36, Asp50, Glu55 and Lys57 and contacts the bases from the minor grove side. The recognition lobe mediates all major grove interactions with the bases. The enzyme in the crystal is bound to the strand with T at the center of the recognition sequence. The crystal structure with calcium ions and DNA mimics the prereactive state. MvaI shows structural similarities to BcnI, which cleaves the related sequence CC/SGG and to MutH enzyme, which is a component of the DNA repair machinery, and nicks one DNA strand instead of making a double-strand break. Oxford University Press 2007-03 2007-03-07 /pmc/articles/PMC1874612/ /pubmed/17344322 http://dx.doi.org/10.1093/nar/gkm064 Text en © 2007 The Author(s) 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 Structural Biology
Kaus-Drobek, Magdalena
Czapinska, Honorata
Sokołowska, Monika
Tamulaitis, Gintautas
Szczepanowski, Roman H.
Urbanke, Claus
Siksnys, Virginijus
Bochtler, Matthias
Restriction endonuclease MvaI is a monomer that recognizes its target sequence asymmetrically
title Restriction endonuclease MvaI is a monomer that recognizes its target sequence asymmetrically
title_full Restriction endonuclease MvaI is a monomer that recognizes its target sequence asymmetrically
title_fullStr Restriction endonuclease MvaI is a monomer that recognizes its target sequence asymmetrically
title_full_unstemmed Restriction endonuclease MvaI is a monomer that recognizes its target sequence asymmetrically
title_short Restriction endonuclease MvaI is a monomer that recognizes its target sequence asymmetrically
title_sort restriction endonuclease mvai is a monomer that recognizes its target sequence asymmetrically
topic Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1874612/
https://www.ncbi.nlm.nih.gov/pubmed/17344322
http://dx.doi.org/10.1093/nar/gkm064
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