DNA topoisomerase IIalpha is required for RNA polymerase II transcription on chromatin templates

Nature. 2001 Sep 27;413(6854):435-8. doi: 10.1038/35096590.

Abstract

In the nucleus of the cell, core RNA polymerase II (pol II) is associated with a large complex called the pol II holoenzyme (holo-pol). Transcription by core pol II in vitro on nucleosomal templates is repressed compared with that on templates of histone-free naked DNA. We found that the transcriptional activity of holo-pol, in contrast to that of core pol II, is not markedly repressed on chromatin templates. We refer to this property of holo-pol as chromatin-dependent coactivation (CDC). Here we show that DNA topoisomerase IIalpha is associated with the holo-pol and is a required component of CDC. Etoposide and ICRF-193, specific inhibitors of topoisomerase II, blocked transcription on chromatin templates, but did not affect transcription on naked templates. Addition of purified topoisomerase IIalpha reconstituted CDC activity in reactions with core pol II. These findings suggest that transcription on chromatin templates results in the accumulation of superhelical tension, making the relaxation activity of topoisomerase II essential for productive RNA synthesis on nucleosomal DNA.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antigens, Neoplasm
  • Cattle
  • Chromatin / metabolism*
  • DNA Topoisomerases, Type II* / physiology*
  • DNA-Binding Proteins
  • Enzyme Inhibitors / pharmacology
  • Etoposide / pharmacology
  • HeLa Cells
  • Holoenzymes / physiology
  • Humans
  • Isoenzymes / antagonists & inhibitors
  • Isoenzymes / physiology*
  • Models, Genetic
  • Piperazines / pharmacology
  • RNA Polymerase II / physiology*
  • Templates, Genetic
  • Topoisomerase II Inhibitors
  • Transcription Factors / metabolism
  • Transcription, Genetic*

Substances

  • Antigens, Neoplasm
  • Chromatin
  • DNA-Binding Proteins
  • Enzyme Inhibitors
  • Holoenzymes
  • Isoenzymes
  • Piperazines
  • Topoisomerase II Inhibitors
  • Transcription Factors
  • 4,4'-(1,2-dimethyl-1,2-ethanediyl)bis-2,6-piperazinedione
  • Etoposide
  • RNA Polymerase II
  • DNA Topoisomerases, Type II