Isolation of genetic suppressor elements, inducing resistance to topoisomerase II-interactive cytotoxic drugs, from human topoisomerase II cDNA
| Autor: | D. P. Suttle, Carolyn R. Zelnick, Igor B. Roninson, William T. Beck, Rama Thimmapaya, Alexander R. Kazarov, Andrei V. Gudkov |
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| Rok vydání: | 1993 |
| Předmět: |
Molecular Sequence Data
Drug Resistance Antineoplastic Agents Biology medicine.disease_cause Polymerase Chain Reaction chemistry.chemical_compound Leukemia Myelogenous Chronic BCR-ABL Positive Complementary DNA Tumor Cells Cultured medicine Humans Topoisomerase II Inhibitors RNA Antisense Cloning Molecular Genes Suppressor Etoposide Gene Library Mutation Multidisciplinary Base Sequence Topoisomerase DNA Neoplasm Molecular biology Antisense RNA DNA Topoisomerases Type II Retroviridae Oligodeoxyribonucleotides chemistry biology.protein TOPO cloning Topoisomerase-II Inhibitor DNA HeLa Cells Plasmids Research Article medicine.drug |
| Zdroj: | Proceedings of the National Academy of Sciences. 90:3231-3235 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.90.8.3231 |
| Popis: | Many cytotoxic anticancer drugs act at topoisomerase II (topo II) by stabilizing cleavable complexes with DNA formed by this enzyme. Several cell lines, selected for resistance to topo II-interactive drugs, show decreased expression or activity of topo II, suggesting that such a decrease may be responsible for drug resistance. In the present study, etoposide resistance was used as the selection strategy to isolate genetic suppressor elements (GSEs) from a retroviral library expressing random fragments of human topo II (alpha form) cDNA. Twelve GSEs were isolated, encoding either peptides corresponding to short segments of the topo II alpha molecule (2.4-6.5% of the protein) or 163- to 220-bp-long antisense RNA sequences. Expression of a GSE encoding antisense RNA led to decreased cellular expression of the topo II alpha protein. Both types of GSE induced resistance to several topo II poisons but not to drugs that do not act at topo II. These results provide direct evidence that inhibition of topo II results in resistance to topo II-interactive drugs, indicate structural domains of topo II capable of independent functional interactions, and demonstrate that expression selection of random fragments constitutes an efficient approach to the generation of GSEs in mammalian cells. |
| Databáze: | OpenAIRE |
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