Document Detail


Isolation of genetic suppressor elements, inducing resistance to topoisomerase II-interactive cytotoxic drugs, from human topoisomerase II cDNA.
MedLine Citation:
PMID:  8386368     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
A V Gudkov; C R Zelnick; A R Kazarov; R Thimmapaya; D P Suttle; W T Beck; I B Roninson
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  90     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1993 Apr 
Date Detail:
Created Date:  1993-05-17     Completed Date:  1993-05-17     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  3231-5     Citation Subset:  IM    
Affiliation:
Department of Genetics, University of Illinois, Chicago 60612-7309.
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MeSH Terms
Descriptor/Qualifier:
Antineoplastic Agents / pharmacology*
Base Sequence
Cloning, Molecular
DNA Topoisomerases, Type II / antagonists & inhibitors*,  genetics*
DNA, Neoplasm / genetics,  isolation & purification
Drug Resistance / genetics*
Etoposide / pharmacology*
Gene Library
Genes, Suppressor*
Hela Cells
Humans
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Molecular Sequence Data
Oligodeoxyribonucleotides
Plasmids
Polymerase Chain Reaction / methods
RNA, Antisense / genetics
Retroviridae / genetics
Tumor Cells, Cultured
Grant Support
ID/Acronym/Agency:
CA39365/CA/NCI NIH HHS; CA40333/CA/NCI NIH HHS; CA56736/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Antineoplastic Agents; 0/DNA, Neoplasm; 0/Oligodeoxyribonucleotides; 0/RNA, Antisense; 33419-42-0/Etoposide; EC 5.99.1.3/DNA Topoisomerases, Type II
Comments/Corrections

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