Newly replicated DNA is associated with DNA topoisomerase II in cultured rat prostatic adenocarcinoma cells

William G. Nelson, Leroy-Fong Liu, Donald S. Coffey

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152 Citations (Scopus)

Abstract

DNA topoisomerases have been proposed to function in a variety of genetic processes in both prokaryotes and eukaryotes1-3. Here, we have assessed the role of DNA topoisomerase II in mammalian DNA replication by determining the proximity of newly synthesized DNA to covalent enzyme-DNA complexes generated by treating cultured rat prostatic adenocarcinoma cells with teniposide. Teniposide (VM-26), an epipodophyllotoxin, is known to interact with mammalian DNA topoisomerase II so as to trap the enzyme in a covalent complex with DNA4-9. We have found that the teniposide-induced trapping of such complexes requires MgCl2, is stimulated by ATP and is inhibited by novobiocin. The formation of covalent complexes seems to be reversible on removal of teniposide. Furthermore, analysis of the covalent complexes formed between 3H-thymidine pulse-labelled DNA and topoisomerase II following teniposide treatment reveals a direct association of the enzyme with nascent DNA fragments. Our results suggest that DNA topoisomerase II may interact with newly replicated daughter DNA molecules near DNA replication forks in mammalian cells.

Original languageEnglish
Pages (from-to)187-189
Number of pages3
JournalNature
Volume322
Issue number6075
DOIs
Publication statusPublished - 1986
Externally publishedYes

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