Spectrum of DNA-platinum adduct recognition by prokaryotic and eukaryotic DNA-dependent RNA polymerases

• Published in: Biochemistry (Easton) Vol. 32; no. 33; pp. 8582 - 8588
• Main Authors: Corda, Yves, Job, Claudette, Anin, Marie France, Leng, Marc, Job, Dominique
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 24.08.1993
• Subjects: Antineoplastic agents; Base Sequence; Biological and medical sciences; DNA - metabolism; DNA-Directed RNA Polymerases - metabolism; Escherichia coli - enzymology; General aspects; Kinetics; Medical sciences; Molecular Sequence Data; Oligodeoxyribonucleotides - chemical synthesis; Oligodeoxyribonucleotides - metabolism; Pharmacology. Drug treatments; Platinum - metabolism; RNA - biosynthesis; Structure-Activity Relationship; Templates, Genetic; Transcription, Genetic; Triticum - enzymology; Antineoplastic agent; Transcription; Enzyme; DNA; RNA polymerase; Mechanism of action; In vitro; Molecular adduct; Platinum II Complexes
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00084a027

Double-stranded DNA oligomers were constructed to evaluate the effect of bifunctional and monofunctional platinum(II) complexes at the level of DNA transcription. They contained a single lesion, which is either a cis-[Pt(NH3)2(d(GpTpG))] intrastrand cross-link, a trans-[Pt(NH3)2(d(GpTpG))] intrastrand cross-link, a cis-[Pt(NH3)2(d(GpC/GpC))] interstrand cross-link, or a (diethylenetriamine)-platinum(II)-dG adduct. The synthetic duplexes were multimerized and then used as templates in dinucleotide-primed reactions catalyzed by prokaryotic or eukaryotic RNA polymerases. Reactions were conducted in the presence of a single triphosphate substrate (single-step addition reaction) or of a combination of triphosphate substrates, permitting elongation of the trinucleotide products to longer RNA chains (productive elongation reaction), respectively. In transcription of the platinated strands, none of the DNA adducts provided an absolute block to formation of a single phosphodiester bond by either Escherichia coli RNA polymerase or wheat germ RNA polymerase II. However, the single-step addition reactions were much more impeded from transcription of bifunctional adduct-containing templates as compared to those containing monofunctional lesions. Productive elongation was irreversibly blocked in transcription of the platinated strand of templates containing a cis-d(G*pTpG*) intrastrand cross-link or a cis-d(G*pC/G*pC) interstrand cross-link. In both cases transcription stopped at the level of the lesion. Termination occurred also several nucleotides before the elongation complexes reached the interstrand cross-link. A substantial amount of the RNA polymerase molecules was able of bypassing the trans-d(G*pTpG*) cross-links. In all the cases single-step addition reactions were enhanced on the template strand complementary to that containing the intrastrand cross-links.