RNA polymerases react differently at d(ApG) and d(GpG) adducts in DNA modified by cis-diamminedichloroplatinum(II).
Identifieur interne : 000374 ( France/Extraction ); précédent : 000373; suivant : 000375RNA polymerases react differently at d(ApG) and d(GpG) adducts in DNA modified by cis-diamminedichloroplatinum(II).
Auteurs : Y. Corda [France] ; M F Anin ; M. Leng ; D. JobSource :
- Biochemistry [ 0006-2960 ] ; 1992.
Descripteurs français
- KwdFr :
- ADN (métabolisme), Adduits à l'ADN, Cisplatine (métabolisme), DNA-directed RNA polymerases (antagonistes et inhibiteurs), DNA-directed RNA polymerases (métabolisme), Dinucléoside phosphates (métabolisme), Données de séquences moléculaires, Désoxyguanosine (métabolisme), Escherichia coli (enzymologie), Hétéroduplexes d'acides nucléiques, Matrices (génétique), Nucléotide désoxyadenylique (métabolisme), Réactifs réticulants, Séquence nucléotidique, Transcription génétique, Triticum (enzymologie).
- MESH :
- antagonistes et inhibiteurs : DNA-directed RNA polymerases.
- enzymologie : Escherichia coli, Triticum.
- métabolisme : ADN, Cisplatine, DNA-directed RNA polymerases, Dinucléoside phosphates, Désoxyguanosine, Nucléotide désoxyadenylique.
- Adduits à l'ADN, Données de séquences moléculaires, Hétéroduplexes d'acides nucléiques, Matrices (génétique), Réactifs réticulants, Séquence nucléotidique, Transcription génétique.
English descriptors
- KwdEn :
- Base Sequence, Cisplatin (metabolism), Cross-Linking Reagents, DNA (metabolism), DNA Adducts, DNA-Directed RNA Polymerases (antagonists & inhibitors), DNA-Directed RNA Polymerases (metabolism), Deoxyadenine Nucleotides (metabolism), Deoxyguanosine (metabolism), Dinucleoside Phosphates (metabolism), Escherichia coli (enzymology), Molecular Sequence Data, Nucleic Acid Heteroduplexes, Templates, Genetic, Transcription, Genetic, Triticum (enzymology).
- MESH :
- chemical , antagonists & inhibitors : DNA-Directed RNA Polymerases.
- chemical , metabolism : Cisplatin, DNA, DNA-Directed RNA Polymerases, Deoxyadenine Nucleotides, Deoxyguanosine, Dinucleoside Phosphates.
- enzymology : Escherichia coli, Triticum.
- Base Sequence, Cross-Linking Reagents, DNA Adducts, Molecular Sequence Data, Nucleic Acid Heteroduplexes, Templates, Genetic, Transcription, Genetic.
Abstract
Two duplexes (20-mers) were constructed containing either a single cis-[Pt(NH3)2[d(GpG)]] or cis-[Pt(NH3)2[d(ApG)]] intrastrand cross-link, the major DNA adducts of the antitumor drug cis-diamminedichloroplatinum(II). These synthetic duplexes were multimerized and the resultant polymers used as templates in single-step addition reactions of condensation of a single nucleoside triphosphate substrate to a dinucleotide primer (abortive elongation reaction) catalyzed by prokaryotic or eukaryotic RNA polymerases. Primer-substrate combinations were selected so as to direct trinucleotide product formation within the platinated bases of the templates. Transcription experiments established that cis-DDP-DNA adducts formed at d(ApG) or d(GpG) sites are not an absolute block to formation of a single phosphodiester bond by either Escherichia coli RNA polymerase or wheat germ RNA polymerase II. Furthermore, the kinetic data indicate that single-step addition reactions are much more impeded at the platinated d(GpG) than at the platinated d(ApG) site and that the mechanisms of inhibition of RNA polymerase activity are different at the two platinated sites. In particular, binding affinity between E. coli RNA polymerase and the d(GpG)-containing platinated template is lowered, as the apparent Km of enzyme for the platinated polymer is increased by a factor of 4-5. In contrast, binding affinity between the RNA polymerase and the d(ApG)-containing template is not affected by modification of the d(ApG) site by cis-diamminedichloroplatinum(II). Similar experiments were carried out with synthetic templates containing the adducts at the d(GpG) sites, in which one of the two platinated dG residues is paired with a dT residue.(ABSTRACT TRUNCATED AT 250 WORDS)
DOI: 10.1021/bi00122a002
PubMed: 1536834
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pubmed:1536834Le document en format XML
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Base Sequence</term>
<term>Cisplatin (metabolism)</term>
<term>Cross-Linking Reagents</term>
<term>DNA (metabolism)</term>
<term>DNA Adducts</term>
<term>DNA-Directed RNA Polymerases (antagonists & inhibitors)</term>
<term>DNA-Directed RNA Polymerases (metabolism)</term>
<term>Deoxyadenine Nucleotides (metabolism)</term>
<term>Deoxyguanosine (metabolism)</term>
<term>Dinucleoside Phosphates (metabolism)</term>
<term>Escherichia coli (enzymology)</term>
<term>Molecular Sequence Data</term>
<term>Nucleic Acid Heteroduplexes</term>
<term>Templates, Genetic</term>
<term>Transcription, Genetic</term>
<term>Triticum (enzymology)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ADN (métabolisme)</term>
<term>Adduits à l'ADN</term>
<term>Cisplatine (métabolisme)</term>
<term>DNA-directed RNA polymerases (antagonistes et inhibiteurs)</term>
<term>DNA-directed RNA polymerases (métabolisme)</term>
<term>Dinucléoside phosphates (métabolisme)</term>
<term>Données de séquences moléculaires</term>
<term>Désoxyguanosine (métabolisme)</term>
<term>Escherichia coli (enzymologie)</term>
<term>Hétéroduplexes d'acides nucléiques</term>
<term>Matrices (génétique)</term>
<term>Nucléotide désoxyadenylique (métabolisme)</term>
<term>Réactifs réticulants</term>
<term>Séquence nucléotidique</term>
<term>Transcription génétique</term>
<term>Triticum (enzymologie)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="antagonists & inhibitors" xml:lang="en"><term>DNA-Directed RNA Polymerases</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Cisplatin</term>
<term>DNA</term>
<term>DNA-Directed RNA Polymerases</term>
<term>Deoxyadenine Nucleotides</term>
<term>Deoxyguanosine</term>
<term>Dinucleoside Phosphates</term>
</keywords>
<keywords scheme="MESH" qualifier="antagonistes et inhibiteurs" xml:lang="fr"><term>DNA-directed RNA polymerases</term>
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<keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Escherichia coli</term>
<term>Triticum</term>
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<keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Escherichia coli</term>
<term>Triticum</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ADN</term>
<term>Cisplatine</term>
<term>DNA-directed RNA polymerases</term>
<term>Dinucléoside phosphates</term>
<term>Désoxyguanosine</term>
<term>Nucléotide désoxyadenylique</term>
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<keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term>
<term>Cross-Linking Reagents</term>
<term>DNA Adducts</term>
<term>Molecular Sequence Data</term>
<term>Nucleic Acid Heteroduplexes</term>
<term>Templates, Genetic</term>
<term>Transcription, Genetic</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Adduits à l'ADN</term>
<term>Données de séquences moléculaires</term>
<term>Hétéroduplexes d'acides nucléiques</term>
<term>Matrices (génétique)</term>
<term>Réactifs réticulants</term>
<term>Séquence nucléotidique</term>
<term>Transcription génétique</term>
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<front><div type="abstract" xml:lang="en">Two duplexes (20-mers) were constructed containing either a single cis-[Pt(NH3)2[d(GpG)]] or cis-[Pt(NH3)2[d(ApG)]] intrastrand cross-link, the major DNA adducts of the antitumor drug cis-diamminedichloroplatinum(II). These synthetic duplexes were multimerized and the resultant polymers used as templates in single-step addition reactions of condensation of a single nucleoside triphosphate substrate to a dinucleotide primer (abortive elongation reaction) catalyzed by prokaryotic or eukaryotic RNA polymerases. Primer-substrate combinations were selected so as to direct trinucleotide product formation within the platinated bases of the templates. Transcription experiments established that cis-DDP-DNA adducts formed at d(ApG) or d(GpG) sites are not an absolute block to formation of a single phosphodiester bond by either Escherichia coli RNA polymerase or wheat germ RNA polymerase II. Furthermore, the kinetic data indicate that single-step addition reactions are much more impeded at the platinated d(GpG) than at the platinated d(ApG) site and that the mechanisms of inhibition of RNA polymerase activity are different at the two platinated sites. In particular, binding affinity between E. coli RNA polymerase and the d(GpG)-containing platinated template is lowered, as the apparent Km of enzyme for the platinated polymer is increased by a factor of 4-5. In contrast, binding affinity between the RNA polymerase and the d(ApG)-containing template is not affected by modification of the d(ApG) site by cis-diamminedichloroplatinum(II). Similar experiments were carried out with synthetic templates containing the adducts at the d(GpG) sites, in which one of the two platinated dG residues is paired with a dT residue.(ABSTRACT TRUNCATED AT 250 WORDS)</div>
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