Chromatin structure, not DNA sequence specificity, is the primary determinant of topoisomerase II sites of action in vivo

A. Udvardy, Paul Schedl

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

In the studies reported here we have used topoisomerase II as a model system for analyzing the factors that determine the sites of action for DNA-binding proteins in vivo. To localize topoisomerase II sites in vivo we used an inhibitor of the purified enzyme, the antitumor drug VM-26. This drug stablizes an intermediate in the catalytic cycle, the cleavable complex, and substantially stimulates DNA cleavage by topoisomerase II. We show that lysis of VM-26 treated tissue culture cells with sodium dodecyl sulfate induces highly specific double-strand breaks in genomic DNA, and we present evidence indicating that these double-strand breaks are generated by topoisomerase II. Using indirect end labeling to map the cleavage products, we have examined the in vivo sites of action of topoisomerase II in the 87A7 heat shock locus, the histone repeat, and a tRNA gene cluster at 90BC. Our analysis reveals that chromatin structure, not sequence specificity, is the primary determinant in topoisomerase II site selection in vivo. We suggest that chromatin organization may provide a general mechanism for generating specificity in a wide range of DNA-protein interactions in vivo.

Original languageEnglish
Pages (from-to)4973-4984
Number of pages12
JournalMolecular and Cellular Biology
Volume11
Issue number10
Publication statusPublished - 1991

Fingerprint

Type II DNA Topoisomerase
Chromatin
Teniposide
DNA Cleavage
DNA
DNA-Binding Proteins
Enzyme Inhibitors
Multigene Family
Transfer RNA
Sodium Dodecyl Sulfate
Antineoplastic Agents
Histones
Shock
Cell Culture Techniques
Hot Temperature
Pharmaceutical Preparations
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Chromatin structure, not DNA sequence specificity, is the primary determinant of topoisomerase II sites of action in vivo. / Udvardy, A.; Schedl, Paul.

In: Molecular and Cellular Biology, Vol. 11, No. 10, 1991, p. 4973-4984.

Research output: Contribution to journalArticle

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