Pseudocomplementary PNAs as selective modifiers of protein activity on duplex DNA: the case of type IIs restriction enzymes
This study evaluates the potential of pseudocomplementary peptide nucleic acids (pcPNAs) for sequence‐specific modification of enzyme activity towards double‐stranded DNA (dsDNA). To this end, we analyze the ability of pcPNA–dsDNA complexes to site‐selectively interfere with the action of four type...
Saved in:
Published in | Nucleic acids research Vol. 31; no. 14; pp. 3929 - 3935 |
---|---|
Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
England
Oxford University Press
15.07.2003
Oxford Publishing Limited (England) |
Subjects | |
Online Access | Get full text |
Cover
Loading…
Summary: | This study evaluates the potential of pseudocomplementary peptide nucleic acids (pcPNAs) for sequence‐specific modification of enzyme activity towards double‐stranded DNA (dsDNA). To this end, we analyze the ability of pcPNA–dsDNA complexes to site‐selectively interfere with the action of four type IIs restriction enzymes. We have found that pcPNA–dsDNA complexes exhibit a different degree of DNA protection against cleaving/nicking activity of various isoschizomeric endonucleases under investigation (PleI, MlyI and N.BstNBI) depending on their type and mutual arrangement of PNA‐binding and enzyme recognition/cleavage sites. We have also found that the pcPNA targeting to closely located PleI or BbsI recognition sites on dsDNA generates in some cases the nicking activity of these DNA cutters. At the same time, MlyI endonuclease, a PleI isoschizomer, does not exhibit any DNA nicking/cleavage activity, being completely blocked by the nearby pcPNA binding. Our results have general implications for effective pcPNA interference with the performance of DNA‐processing proteins, thus being important for prospective applications of pcPNAs. |
---|---|
Bibliography: | ark:/67375/HXZ-77J827M2-6 Received April 18, 2003; Revised and Accepted May 13, 2003 istex:F4DC66B13CE0A920C3952C093C4BC9035AA81F21 local:gkg450 To whom correspondence should be addressed. Tel: +1 617 353 8498; Fax: +1 617 353 8501; Email: mfk@bu.edu The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 To whom correspondence should be addressed. Tel: +1 617 353 8498; Fax: +1 617 353 8501; Email: mfk@bu.edu The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors |
ISSN: | 0305-1048 1362-4962 1362-4962 |
DOI: | 10.1093/nar/gkg450 |