Linked Life Data

10547285

Source:http://linkedlifedata.com/resource/pubmed/id/10547285

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2000-1-11
pubmed:databankReference
pubmed:abstractText
The HaeIV restriction endonuclease (ENase) belongs to a distinct class of ENases, characterized by its ability to cleave double-stranded DNA on both sides of its recognition sequence, excising a short DNA fragment that includes the recognition sequence. The gene encoding the HaeIV ENase was cloned from Haemophilus aegyptius into pUC19 using a previously described system that does not need the knowledge that a particular ENase is produced by a bacterial strain. DNA sequence analysis of the insert contained on this plasmid identified a single open reading frame (ORF), with the predicted protein having an apparent molecular mass of approximately 110 kDa. The protein encoded by this ORF was purified to homogeneity from Escherichia coli strain ER1944 carrying the haeIVRM gene on a recombinant plasmid under the control of the inducible ara promoter. The protein possessed both ENase and methyltransferase (MTase) activities. Amino acid sequence analysis was able to identify several conserved motifs found in DNA MTases, located in the middle of the protein. The enzyme recognizes the interrupted palindromic sequence 5' GAPyNNNNNPuTC 3', cleaving double-stranded DNA on both strands upstream and downstream of the recognition sequence, releasing an approximately 33 bp fragment. The ENase possessed an absolute requirement only for Mg(+2). ATP had no influence on ENase or MTase activities. The ENase made the first strand cleavage randomly on either side of the recognition sequence, but the second cleavage occurred more slowly. The MTase activity modified symmetrically located adenine residues on both strands within the recognition sequence yielding N6-methyl adenine. Furthermore, the MTase was active as a dimer.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0022-2836
pubmed:author
pubmed:copyrightInfo
Copyright 1999 Academic Press.
pubmed:issnType
Print
pubmed:day
12
pubmed:volume
293
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1055-65
pubmed:dateRevised
2010-1-20
pubmed:meshHeading
pubmed-meshheading:10547285-Adenosine Triphosphate, pubmed-meshheading:10547285-Amino Acid Sequence, pubmed-meshheading:10547285-Base Sequence, pubmed-meshheading:10547285-Cloning, Molecular, pubmed-meshheading:10547285-Conserved Sequence, pubmed-meshheading:10547285-DNA, pubmed-meshheading:10547285-DNA Methylation, pubmed-meshheading:10547285-DNA Modification Methylases, pubmed-meshheading:10547285-DNA Restriction Enzymes, pubmed-meshheading:10547285-Dimerization, pubmed-meshheading:10547285-Genes, Bacterial, pubmed-meshheading:10547285-Haemophilus, pubmed-meshheading:10547285-Kinetics, pubmed-meshheading:10547285-Magnesium, pubmed-meshheading:10547285-Molecular Sequence Data, pubmed-meshheading:10547285-Molecular Weight, pubmed-meshheading:10547285-Open Reading Frames, pubmed-meshheading:10547285-Peptides, pubmed-meshheading:10547285-Recombinant Proteins, pubmed-meshheading:10547285-S-Adenosylmethionine, pubmed-meshheading:10547285-Sequence Alignment, pubmed-meshheading:10547285-Substrate Specificity
pubmed:year
1999
pubmed:articleTitle
The HaeIV restriction modification system of Haemophilus aegyptius is encoded by a single polypeptide.
pubmed:affiliation
Institute of Microbiology, University of Warsaw, Nowy Swiat 67, Warsaw, 00-046, Poland. ANPIEK@PLEARN.EDU.PL
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't