RNA Dependent DNA Replication Fidelity of HIV-1 Reverse Transcriptase: Evidence of Discrimination between DNA and RNA Substrates†
Identifieur interne : 000B19 ( Istex/Curation ); précédent : 000B18; suivant : 000B20RNA Dependent DNA Replication Fidelity of HIV-1 Reverse Transcriptase: Evidence of Discrimination between DNA and RNA Substrates†
Auteurs : Stephen G. Kerr [États-Unis] ; Karen S. Anderson [États-Unis]Source :
- Biochemistry [ 0006-2960 ] ; 1997.
Abstract
The RNA dependent DNA replication fidelity of HIV-1 reverse transcriptase has been investigated using pre-steady-state kinetics under single turnover conditions. In contrast to previous estimates of low replication fidelity of HIV-1 reverse transcriptase, the present study finds the enzyme to be more highly discriminating when an RNA/DNA template−primer is employed as compared with the corresponding DNA/DNA template−primer. The basis of this selectivity is due to extremely slow polymerization kinetics for incorporation of an incorrect deoxynucleotide. The maximum rates for misincorporation (kpol) of dGTP, dCTP, and dTTP opposite a template uridine were 0.2, 0.03, and 0.003 s-1, respectively. The equilibrium dissociation constants (Kd) for the incorrect nucleotide opposite a template uridine were 1.0, 1.1, and 0.7 mM for dGTP, dCTP, and dTTP, respectively. These kinetic values provide fidelity estimates of 26 000 for discrimination against dGTP, 176 000 for dCTP, and 1 × 106 for dTTP misincorporation at this position. Similar observations were obtained when incorrect nucleotide misincorporation was examined opposite a template adenine. Thus in a direct comparison of RNA/DNA and DNA/DNA template−primer substrates, HIV-1 RT exhibits approximately a 10−60-fold increase in fidelity. This study augments our current understanding of the similarities and differences of catalytic activity of HIV-1 reverse transcriptase using RNA and DNA substrates. Moreover, these studies lend further support for a model for nucleotide incorporation by HIV-1 reverse transcriptase involving a two-step binding mechanism governed by a rate-limiting conformational change for correct incorporation.
Url:
DOI: 10.1021/bi971385+
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: Pour aller vers cette notice dans l'étape Curation :000B19
Links to Exploration step
ISTEX:CABC69F131E4A9F0B6211788915355DF3130DEC7Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>RNA Dependent DNA Replication Fidelity of HIV-1 Reverse Transcriptase: Evidence of Discrimination between DNA and RNA Substrates†</title><author><name sortKey="Kerr, Stephen G" sort="Kerr, Stephen G" uniqKey="Kerr S" first="Stephen G." last="Kerr">Stephen G. Kerr</name><affiliation wicri:level="2"><mods:affiliation>Department of Pharmacology, 333 Cedar Street, Yale University School of Medicine, New Haven, Connecticut 06520-8066</mods:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">Connecticut</region></placeName><wicri:cityArea>Department of Pharmacology, 333 Cedar Street, Yale University School of Medicine, New Haven</wicri:cityArea></affiliation><affiliation wicri:level="2"><mods:affiliation> Present address: Massachusetts College ofPharmacy & AlliedHealth Sciences, 179 Longwood Ave., Boston, MA 02115.</mods:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">Massachusetts</region></placeName><wicri:cityArea> Present address: Massachusetts College ofPharmacy & AlliedHealth Sciences, 179 Longwood Ave., Boston</wicri:cityArea></affiliation></author><author><name sortKey="Anderson, Karen S" sort="Anderson, Karen S" uniqKey="Anderson K" first="Karen S." last="Anderson">Karen S. Anderson</name><affiliation wicri:level="2"><mods:affiliation>Department of Pharmacology, 333 Cedar Street, Yale University School of Medicine, New Haven, Connecticut 06520-8066</mods:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">Connecticut</region></placeName><wicri:cityArea>Department of Pharmacology, 333 Cedar Street, Yale University School of Medicine, New Haven</wicri:cityArea></affiliation><affiliation wicri:level="1"><mods:affiliation> Author to whom correspondence should be addressed.Telephone:(203)-785-4526. Fax: (203)-785-7670. email:karen.anderson@yale.edu.</mods:affiliation><country wicri:rule="url">États-Unis</country></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:CABC69F131E4A9F0B6211788915355DF3130DEC7</idno><date when="1997" year="1997">1997</date><idno type="doi">10.1021/bi971385+</idno><idno type="url">https://api.istex.fr/ark:/67375/TPS-67D2GZM8-Q/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000B19</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000B19</idno><idno type="wicri:Area/Istex/Curation">000B19</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main">RNA Dependent DNA Replication Fidelity of HIV-1 Reverse Transcriptase:
Evidence of Discrimination between DNA and RNA Substrates<ref type="bib" target="#bi9713851AF2"><hi rend="superscript">†</hi></ref></title><author><name sortKey="Kerr, Stephen G" sort="Kerr, Stephen G" uniqKey="Kerr S" first="Stephen G." last="Kerr">Stephen G. Kerr</name><affiliation wicri:level="2"><mods:affiliation>Department of Pharmacology, 333 Cedar Street, Yale University School of Medicine, New Haven, Connecticut 06520-8066</mods:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">Connecticut</region></placeName><wicri:cityArea>Department of Pharmacology, 333 Cedar Street, Yale University School of Medicine, New Haven</wicri:cityArea></affiliation><affiliation wicri:level="2"><mods:affiliation> Present address: Massachusetts College ofPharmacy & AlliedHealth Sciences, 179 Longwood Ave., Boston, MA 02115.</mods:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">Massachusetts</region></placeName><wicri:cityArea> Present address: Massachusetts College ofPharmacy & AlliedHealth Sciences, 179 Longwood Ave., Boston</wicri:cityArea></affiliation></author><author><name sortKey="Anderson, Karen S" sort="Anderson, Karen S" uniqKey="Anderson K" first="Karen S." last="Anderson">Karen S. Anderson</name><affiliation wicri:level="2"><mods:affiliation>Department of Pharmacology, 333 Cedar Street, Yale University School of Medicine, New Haven, Connecticut 06520-8066</mods:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">Connecticut</region></placeName><wicri:cityArea>Department of Pharmacology, 333 Cedar Street, Yale University School of Medicine, New Haven</wicri:cityArea></affiliation><affiliation wicri:level="1"><mods:affiliation> Author to whom correspondence should be addressed.Telephone:(203)-785-4526. Fax: (203)-785-7670. email:karen.anderson@yale.edu.</mods:affiliation><country wicri:rule="url">États-Unis</country></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Biochemistry</title><title level="j" type="abbrev">Biochemistry</title><idno type="ISSN">0006-2960</idno><idno type="eISSN">1520-4995</idno><imprint><publisher>American Chemical Society</publisher><date type="e-published">1997</date><date type="published">1997</date><biblScope unit="vol">36</biblScope><biblScope unit="issue">46</biblScope><biblScope unit="page" from="14056">14056</biblScope><biblScope unit="page" to="14063">14063</biblScope></imprint><idno type="ISSN">0006-2960</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0006-2960</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract">The RNA dependent DNA replication fidelity of HIV-1 reverse transcriptase has been investigated using pre-steady-state kinetics under single turnover conditions. In contrast to previous estimates of low replication fidelity of HIV-1 reverse transcriptase, the present study finds the enzyme to be more highly discriminating when an RNA/DNA template−primer is employed as compared with the corresponding DNA/DNA template−primer. The basis of this selectivity is due to extremely slow polymerization kinetics for incorporation of an incorrect deoxynucleotide. The maximum rates for misincorporation (kpol) of dGTP, dCTP, and dTTP opposite a template uridine were 0.2, 0.03, and 0.003 s-1, respectively. The equilibrium dissociation constants (Kd) for the incorrect nucleotide opposite a template uridine were 1.0, 1.1, and 0.7 mM for dGTP, dCTP, and dTTP, respectively. These kinetic values provide fidelity estimates of 26 000 for discrimination against dGTP, 176 000 for dCTP, and 1 × 106 for dTTP misincorporation at this position. Similar observations were obtained when incorrect nucleotide misincorporation was examined opposite a template adenine. Thus in a direct comparison of RNA/DNA and DNA/DNA template−primer substrates, HIV-1 RT exhibits approximately a 10−60-fold increase in fidelity. This study augments our current understanding of the similarities and differences of catalytic activity of HIV-1 reverse transcriptase using RNA and DNA substrates. Moreover, these studies lend further support for a model for nucleotide incorporation by HIV-1 reverse transcriptase involving a two-step binding mechanism governed by a rate-limiting conformational change for correct incorporation.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Istex/Curation
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000B19 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Istex/Curation/biblio.hfd -nk 000B19 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Istex
|étape= Curation
|type= RBID
|clé= ISTEX:CABC69F131E4A9F0B6211788915355DF3130DEC7
|texte= RNA Dependent DNA Replication Fidelity of HIV-1 Reverse Transcriptase: Evidence of Discrimination between DNA and RNA Substrates†
}}
| This area was generated with Dilib version V0.6.33. |  |