Crystal structures of 2-acetylaminofluorene and 2-aminofluorene in complex with T7 DNA polymerase reveal mechanisms of mutagenesis
| Autor: | Charles C. Richardson, Tom Ellenberger, Shuchismita Dutta, Ying Li, Louis J. Romano, Leonid Dzantiev, Donald E. Johnson |
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| Rok vydání: | 2004 |
| Předmět: |
DNA Replication
Models Molecular Macromolecular Substances Stereochemistry DNA polymerase Molecular Conformation DNA-Directed DNA Polymerase Crystallography X-Ray DNA polymerase delta chemistry.chemical_compound polycyclic compounds Polymerase Fluorenes Multidisciplinary DNA clamp Base Sequence biology Chemistry Mutagenesis DNA replication T7 DNA polymerase DNA 2-Acetylaminofluorene Biological Sciences Biochemistry Carcinogens biology.protein |
| Zdroj: | Proceedings of the National Academy of Sciences. 101:16186-16191 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.0406516101 |
| Popis: | The carcinogen 2-acetylaminofluorene forms two major DNA adducts: N -(2′-deoxyguanosin-8-yl)-2-acetylaminofluorene (dG-AAF) and its deacetylated derivative, N -(2′-deoxyguanosin-8-yl)-2-aminofluorene (dG-AF). Although the dG-AAF and dG-AF adducts are distinguished only by the presence or absence of an acetyl group, they have profoundly different effects on DNA replication. dG-AAF poses a strong block to DNA synthesis and primarily induces frameshift mutations in bacteria, resulting in the loss of one or two nucleotides during replication past the lesion. dG-AF is less toxic and more easily bypassed by DNA polymerases, albeit with an increased frequency of misincorporation opposite the lesion, primarily resulting in G → T transversions. We present three crystal structures of bacteriophage T7 DNA polymerase replication complexes, one with dG-AAF in the templating position and two others with dG-AF in the templating position. Our crystallographic data suggest why a dG-AAF adduct blocks replication more strongly than does a dG-AF adduct and provide a possible explanation for frameshift mutagenesis during replication bypass of a dG-AAF adduct. The dG-AAF nucleoside adopts a syn conformation that facilitates the intercalation of its fluorene ring into a hydrophobic pocket on the surface of the fingers subdomain and locks the fingers in an open, inactive conformation. In contrast, the dG-AF base at the templating position is not well defined by the electron density, consistent with weak binding to the polymerase and a possible interchange of this adduct between the syn and anti conformations. |
| Databáze: | OpenAIRE |
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