Publications
2003
Wilhelm F X, Wilhelm M, Gabriel A
Extension and cleavage of the polypurine tract plus-strand primer by Ty1 reverse transcriptase Article de journal
Dans: J Biol Chem, vol. 278, no. 48, p. 47678-47684, 2003, ISBN: 14500728, (0021-9258 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Base Sequence DNA/chemistry DNA Primers DNA Replication Models, Calf Thymus/chemistry Support, Genetic, Genetic Molecular Sequence Data Purines/*chemistry RNA/chemistry RNA, Messenger/metabolism RNA, Non-U.S. Gov't Support, P.H.S. Templates, U.S. Gov't, Unité ARN, Viral RNA-Directed DNA Polymerase/*chemistry Recombinant Proteins/chemistry Retroelements/*genetics Ribonuclease H
@article{,
title = {Extension and cleavage of the polypurine tract plus-strand primer by Ty1 reverse transcriptase},
author = {F X Wilhelm and M Wilhelm and A Gabriel},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14500728},
isbn = {14500728},
year = {2003},
date = {2003-01-01},
journal = {J Biol Chem},
volume = {278},
number = {48},
pages = {47678-47684},
abstract = {Using hybrid RNA/DNA substrates containing the polypurine tract (PPT) plus-strand primer, we have examined the interaction between the Ty1 reverse transcriptase (RT) and the plus-strand initiation complex. We show here that, although the PPT sequence is relatively resistant to RNase H cleavage, it can be cleaved internally by the polymerase-independent RNase H activity of Ty1 RT. Alternatively, this PPT can be used to initiate plus-strand DNA synthesis. We demonstrate that cleavage at the PPT/DNA junction occurs only after at least 9 nucleotides are extended. Cleavage leaves a nick between the RNA primer and the nascent plus-strand DNA. We show that Ty1 RT has a strand displacement activity beyond a gap but that the PPT is not efficiently re-utilized in vitro for another round of DNA synthesis after a first plus-strand DNA has been synthesized and cleaved at the PPT/U3 junction.},
note = {0021-9258
Journal Article},
keywords = {Base Sequence DNA/chemistry DNA Primers DNA Replication Models, Calf Thymus/chemistry Support, Genetic, Genetic Molecular Sequence Data Purines/*chemistry RNA/chemistry RNA, Messenger/metabolism RNA, Non-U.S. Gov't Support, P.H.S. Templates, U.S. Gov't, Unité ARN, Viral RNA-Directed DNA Polymerase/*chemistry Recombinant Proteins/chemistry Retroelements/*genetics Ribonuclease H},
pubstate = {published},
tppubtype = {article}
}
Using hybrid RNA/DNA substrates containing the polypurine tract (PPT) plus-strand primer, we have examined the interaction between the Ty1 reverse transcriptase (RT) and the plus-strand initiation complex. We show here that, although the PPT sequence is relatively resistant to RNase H cleavage, it can be cleaved internally by the polymerase-independent RNase H activity of Ty1 RT. Alternatively, this PPT can be used to initiate plus-strand DNA synthesis. We demonstrate that cleavage at the PPT/DNA junction occurs only after at least 9 nucleotides are extended. Cleavage leaves a nick between the RNA primer and the nascent plus-strand DNA. We show that Ty1 RT has a strand displacement activity beyond a gap but that the PPT is not efficiently re-utilized in vitro for another round of DNA synthesis after a first plus-strand DNA has been synthesized and cleaved at the PPT/U3 junction.