Nucleic Acids Research Advance Access originally published online on March 7, 2007
Nucleic Acids Research 2007 35(6):2035-2046; doi:10.1093/nar/gkm064
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Nucleic Acids Research, 2007, Vol. 35, No. 6 2035-2046
© 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.
Structural Biology |
Restriction endonuclease MvaI is a monomer that recognizes its target sequence asymmetrically
owska1,21International Institute of Molecular and Cell Biology, ul. Trojdena 4, 02-109 Warsaw, Poland, 2Max-Planck-Institute for Molecular Cell Biology and Genetics, Pfotenhauerstr. 108, 01309 Dresden, Germany, 3Institute of Biotechnology, Graiciuno 8, LT-02241, Vilnius, Lithuania and 4Medizinische Hochschule, Abteilung Strukturanalyse OE 8830, Carl Neuberg Str. 1, 30625 Hannover, Germany
*To whom correspondence should be addressed. Tel: 0048 22 5970732; Fax: 0048 22 5970715; Email: MBochtler{at}iimcb.gov.pl
Correspondence may also be addressed to Virginijus Siksnys. Tel: 00370 5 2602108; Fax: 00370 5 2602116; Email: siksyns{at}ibt.lt
Received December 15, 2006. Revised January 19, 2007. Accepted January 19, 2007.
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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