Linked Life Data

12589753

Source:http://linkedlifedata.com/resource/pubmed/id/12589753

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2003-2-18
pubmed:abstractText
BfiI is a novel type IIs restriction endonuclease that, unlike all other restriction enzymes characterised to date, cleaves DNA in the absence of Mg(2+). The amino acid sequence of the N-terminal part of BfiI has some similarities to Nuc of Salmonella typhimurium, an EDTA-resistant nuclease akin to phospholipase D. The dimeric form of Nuc contains a single active site composed of residues from both subunits. To examine the roles of the amino acid residues of BfiI that align with the catalytic residues in Nuc, a set of alanine replacement mutants was generated by site-directed mutagenesis. The mutationally altered forms of BfiI were all catalytically inactive but were still able to bind DNA specifically. The active site of BfiI is thus likely to be similar to that of Nuc. BfiI was also found by gel-filtration to be a dimer in solution. Both gel-shift and pull-down assays indicated that the dimeric form of BfiI binds two copies of its recognition sequence. In reactions on plasmids with either one or two copies of its recognition sequence, BfiI cleaved the DNA with two sites more rapidly than that with one site. Yet, when bound to two copies of its recognition sequence, the BfiI dimer cleaved only one phosphodiester bond at a time. The dimer thus seems to contain two DNA-binding domains but only one active site.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0022-2836
pubmed:author
pubmed:issnType
Print
pubmed:day
28
pubmed:volume
326
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1051-64
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:12589753-Amino Acid Sequence, pubmed-meshheading:12589753-Bacterial Proteins, pubmed-meshheading:12589753-Binding Sites, pubmed-meshheading:12589753-Catalytic Domain, pubmed-meshheading:12589753-DNA, pubmed-meshheading:12589753-Deoxyribonucleases, Type II Site-Specific, pubmed-meshheading:12589753-Dimerization, pubmed-meshheading:12589753-Endonucleases, pubmed-meshheading:12589753-Micrococcal Nuclease, pubmed-meshheading:12589753-Models, Molecular, pubmed-meshheading:12589753-Molecular Sequence Data, pubmed-meshheading:12589753-Mutagenesis, Site-Directed, pubmed-meshheading:12589753-Oligonucleotides, pubmed-meshheading:12589753-Plasmids, pubmed-meshheading:12589753-Protein Binding, pubmed-meshheading:12589753-Protein Structure, Quaternary, pubmed-meshheading:12589753-Sequence Alignment
pubmed:year
2003
pubmed:articleTitle
The metal-independent type IIs restriction enzyme BfiI is a dimer that binds two DNA sites but has only one catalytic centre.
pubmed:affiliation
Institute of Biotechnology, Graiciuno 8, Vilnius 2028, Lithuania.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't