Bizelesin, a Bifunctional Cyclopropylpyrroloindole Alkylating Agent, Inhibits Simian Virus 40 Replication in Trans by Induction of an Inhibitor
| Autor: | Mary M. McHugh, Mary H. Walsh-O'Beirne, Jen-Sing Liu, Shu-Ru Kuo, Thomas Melendy, Terry A. Beerman |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
DNA Replication
Alkylating Agents Hot Temperature Indoles viruses Simian virus 40 Virus Replication Antiviral Agents Biochemistry Virus Cell Line HeLa DNA Adducts Duocarmycins chemistry.chemical_compound Chlorocebus aethiops Animals Humans Urea Electrophoresis Gel Two-Dimensional Replicon Cell-Free System Dose-Response Relationship Drug biology DNA replication biology.organism_classification Virology Molecular biology In vitro chemistry DNA Viral Agarose gel electrophoresis Trans-Activators Trans-acting DNA DNA Damage HeLa Cells |
| Zdroj: | Biochemistry. 38:11508-11515 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi990598r |
| Popis: | Bizelesin, a bifunctional DNA minor groove alkylating agent, inhibits both cellular and viral (SV40) DNA replication in whole cells. Bizelesin inhibition of SV40 DNA replication was analyzed in SV40-infected cells, using two-dimensional (2D) neutral agarose gel electrophoresis, and in a cell-free SV40 DNA replication assay. Within 1 h of bizelesin addition to infected cells, a similar rapid decrease in both the level of SV40 replication intermediates and replication activity was observed, indicating inhibition of initiation of SV40 DNA replication. However, prolonged bizelesin treatment (>/=2 h) was associated with a reduced extent of elongation of SV40 replicons, as well as the appearance on 2D gels of intense spots, suggestive of replication pause sites. Inhibition of elongation and induction of replication pause sites may result from the formation of bizelesin covalent bonds on replicating SV40 molecules. The level of in vitro replication of SV40 DNA also was reduced when extracts from bizelesin-treated HeLa cells were used. This effect was not dependent upon the formation of bizelesin covalent bonds with the template DNA. Mixing experiments, using extracts from control and bizelesin-treated cells, indicated that reduced DNA replication competence was due to the presence of a trans-acting DNA replication inhibitor, rather than to decreased levels or inactivation of essential replication factor(s). |
| Databáze: | OpenAIRE |
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