Structures of the rare-cutting restriction endonuclease NotI reveal a unique metal binding fold involved in DNA binding.

Lambert, Abigail R and Sussman, Django and Shen, Betty and Maunus, Robert and Nix, Jay and Samuelson, James and Xu, Shuang-Yong and Stoddard, Barry L (2008) Structures of the rare-cutting restriction endonuclease NotI reveal a unique metal binding fold involved in DNA binding. Structure (London, England : 1993), 16 (4). pp. 558-569. ISSN 0969-2126

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Abstract

The structure of the rare-cutting restriction endonuclease NotI, which recognizes the 8 bp target 5'-GCGGCCGC-3', has been solved with and without bound DNA. Because of its specificity (recognizing a site that occurs once per 65 kb), NotI is used to generate large genomic fragments and to map DNA methylation status. NotI contains a unique metal binding fold, found in a variety of putative endonucleases, occupied by an iron atom coordinated within a tetrahedral Cys4 motif. This domain positions nearby protein elements for DNA recognition, and serves a structural role. While recognition of the central six base pairs of the target is accomplished via a saturated hydrogen bond network typical of restriction enzymes, the most peripheral base pairs are engaged in a single direct contact in the major groove, reflecting reduced pressure to recognize those positions. NotI may represent an evolutionary intermediate between mobile endonucleases (which recognize longer target sites) and canonical restriction endonucleases.

Item Type:	Article or Abstract
DOI:	10.1016/j.str.2008.01.017
PubMed ID:	18400177
NIHMSID:	NIHMS46475
PMCID:	PMC2390919
Grant Numbers:	T32 GM07270, R01 GM49857
Keywords or MeSH Headings:	Amino Acid Sequence; Binding Sites; Crystallography, X-Ray; DNA/chemistry; Deoxyribonucleases, Type II Site-Specific/chemistry; Iron/chemistry; Models, Molecular; Molecular Sequence Data; Protein Binding; Protein Folding; Protein Structure, Tertiary; Sequence Homology, Amino Acid; Substrate Specificity;
Subjects:	Molecules > Proteins > Enzymes Molecules > Molecular structure
Depositing User:	Library Staff
Date Deposited:	30 Jul 2008 05:01
Last Modified:	14 Feb 2012 14:42
URI:	http://authors.fhcrc.org/id/eprint/15

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