Determination of DNA guanine sites forming photo-adducts with Ru(II)-labeled oligonucleotides; DNA polymerase inhibition by the resulting photo-crosslinking
| Autor: | Olivier Lentzen, Stephan Schumm, Eric Defrancq, Jean-François Constant, Cécile Moucheron, Andrée Kirsch-De Mesmaeker, David García-Fresnadillo, Pascal Dumy |
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| Rok vydání: | 2003 |
| Předmět: |
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Photochemistry Guanine DNA polymerase Stereochemistry Antineoplastic Agents DNA-Directed DNA Polymerase Biochemistry Ruthenium Adduct Inorganic Chemistry DNA Adducts chemistry.chemical_compound Electron transfer DNA Polymerase beta Guanidine biology Oligonucleotide hemic and immune systems respiratory system DNA Polymerase I Cross-Linking Reagents Oligodeoxyribonucleotides chemistry Duplex (building) Molecular Probes biology.protein DNA Conjugate |
| Zdroj: | JBIC Journal of Biological Inorganic Chemistry. 9:100-108 |
| ISSN: | 1432-1327 0949-8257 |
| DOI: | 10.1007/s00775-003-0502-3 |
| Popis: | The influence of the distance between the an- choring site of the tethered (Ru(TAP)2dip) 2+ complex (TAP=1,4,5,8-tetraazaphenanthrene; dip=4,7-diphe- nyl-1,10-phenanthroline) on a probe sequence and the guanines of the complementary target strand was stud- ied by (1) the luminescence quenching of the complex (by electron transfer) and (2) the oligodeoxyribonu- cleotide adduct (ODN adduct) formation which results in photo-crosslinking of the two strands. Moving the guanine moieties away from the complex induces an important decrease of the efficiency of both processes, but clearly affects the ODN adduct formation more specifically than the quenching process. From these re- sults, we determined the positions of the guanine bases in the duplex ODN that are able to form a photo-adduct with the tethered complex. We also examined the pos- sible competition between a long-range hole migration in the duplex ODN and the formation of a photo-adduct by using a sequence labeled with the complex at the 5¢- phosphate end. Such a hole migration appears to be inefficient as compared to the ODN adduct formation. Finally, we studied the influence of the photo-cross- linking on the function of two different DNA polyme- rases. A 17-mer Ru(II)-labeled ODN was hybridized to its complementary sequence located on the 5¢-side of a 40-mer matrix. After illumination, the elongation of a 13-mer DNA primer hybridized to the 3¢-extremity of the same matrix was stopped at a position correspond- ing to the formation of the ODN adduct. |
| Databáze: | OpenAIRE |
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