Probing structure/function relationships of HIV-1 reverse transcriptase with styrene oxide N2-guanine adducts.
Identifieur interne : 002731 ( PubMed/Curation ); précédent : 002730; suivant : 002732Probing structure/function relationships of HIV-1 reverse transcriptase with styrene oxide N2-guanine adducts.
Auteurs : E. Forgacs [États-Unis] ; G. Latham ; W A Beard ; R. Prasad ; K. Bebenek ; T A Kunkel ; S H Wilson ; R S LloydSource :
- The Journal of biological chemistry [ 0021-9258 ] ; 1997.
Descripteurs français
- KwdFr :
- ADN (métabolisme), Adduits à l'ADN (métabolisme), Composés époxy (métabolisme), Conformation d'acide nucléique, Données de séquences moléculaires, Guanine (métabolisme), Isomérie, Matrices (génétique), Modèles moléculaires, Réplication de l'ADN, Séquence nucléotidique, Transcriptase inverse du VIH (génétique), Transcriptase inverse du VIH (métabolisme).
- MESH :
English descriptors
- KwdEn :
- Base Sequence, DNA (metabolism), DNA Adducts (metabolism), DNA Replication, Epoxy Compounds (metabolism), Guanine (metabolism), HIV Reverse Transcriptase (genetics), HIV Reverse Transcriptase (metabolism), Isomerism, Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Templates, Genetic.
- MESH :
- chemical , genetics : HIV Reverse Transcriptase.
- chemical , metabolism : DNA, DNA Adducts, Epoxy Compounds, Guanine, HIV Reverse Transcriptase.
- Base Sequence, DNA Replication, Isomerism, Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Templates, Genetic.
Abstract
Details of the interactions between the human immunodeficiency virus (HIV-1) reverse transcriptase and substrate DNA were probed both by introducing site-specific and stereospecific modifications into DNA and by altering the structure of potential critical residues in the polymerase. Unadducted 11-mer DNAs and 11-mer DNAs containing R and S enantiomers of styrene oxide at N2-guanine were ligated with two additional oligonucleotides to create 63-mers that served as templates for HIV-1 reverse transcriptase replication. Oligonucleotides that primed synthesis 5 bases 3' to the adducts could be extended up to 1 base 3' and opposite the lesion. However, when the positions of the 3'-OH of the priming oligonucleotides were placed 1, 2, 3, 4, 5, and 6 bases downstream of the styrene oxide guanine adducts, replication was initiated, only to be blocked after incorporating 4, 5, 6, and 7 bases beyond the lesion. The sites of this adduct-induced termination corresponded to the position of the DNA where alpha-helix H makes contact with the DNA minor groove, 3-5 bases upstream of the growing 3' end. In addition, mutants of the polymerase in alpha-helix H (W266A and G262A) alter the termination probabilities caused by these DNA adducts, suggesting that alpha-helix H is a sensitive monitor of modifications in the minor groove of newly synthesized template-primer DNA several bases distal to the 3'-OH.
DOI: 10.1074/jbc.272.13.8525
PubMed: 9079681
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pubmed:9079681Le document en format XML
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<term>Molecular Sequence Data</term>
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<term>Matrices (génétique)</term>
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<front><div type="abstract" xml:lang="en">Details of the interactions between the human immunodeficiency virus (HIV-1) reverse transcriptase and substrate DNA were probed both by introducing site-specific and stereospecific modifications into DNA and by altering the structure of potential critical residues in the polymerase. Unadducted 11-mer DNAs and 11-mer DNAs containing R and S enantiomers of styrene oxide at N2-guanine were ligated with two additional oligonucleotides to create 63-mers that served as templates for HIV-1 reverse transcriptase replication. Oligonucleotides that primed synthesis 5 bases 3' to the adducts could be extended up to 1 base 3' and opposite the lesion. However, when the positions of the 3'-OH of the priming oligonucleotides were placed 1, 2, 3, 4, 5, and 6 bases downstream of the styrene oxide guanine adducts, replication was initiated, only to be blocked after incorporating 4, 5, 6, and 7 bases beyond the lesion. The sites of this adduct-induced termination corresponded to the position of the DNA where alpha-helix H makes contact with the DNA minor groove, 3-5 bases upstream of the growing 3' end. In addition, mutants of the polymerase in alpha-helix H (W266A and G262A) alter the termination probabilities caused by these DNA adducts, suggesting that alpha-helix H is a sensitive monitor of modifications in the minor groove of newly synthesized template-primer DNA several bases distal to the 3'-OH.</div>
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