BRCA1 interaction with RNA polymerase II reveals a role for hRPB2 and hRPB10alpha in activated transcription

Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3148-53. doi: 10.1073/pnas.97.7.3148.

Abstract

The functions of most of the 12 subunits of the RNA polymerase II (Pol II) enzyme are unknown. In this study, we demonstrate that two of the subunits, hRPB2 and hRPB10alpha, mediate the regulated stimulation of transcription. We find that the transcriptional coactivator BRCA1 interacts directly with the core Pol II complex in vitro. We tested whether single subunits from Pol II would compete with the intact Pol II complex to inhibit transcription stimulated by BRCA1. Excess purified Pol II subunits hRPB2 or hRPB10alpha blocked BRCA1- and VP16-dependent transcriptional activation in vitro with minimal effect on basal transcription. No other Pol II subunits tested inhibited activated transcription in these assays. Furthermore, hRPB10alpha, but not hRPB2, blocked Sp1-dependent activation.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • BRCA1 Protein / metabolism*
  • Cloning, Molecular
  • Humans
  • Molecular Sequence Data
  • Protein Binding
  • RNA Polymerase II / genetics
  • RNA Polymerase II / metabolism*
  • RNA Polymerase II / physiology
  • Recombinant Proteins / metabolism
  • Sp1 Transcription Factor / metabolism
  • Trans-Activators / metabolism
  • Transcriptional Activation / physiology*

Substances

  • BRCA1 Protein
  • Gal-VP16
  • Recombinant Proteins
  • Sp1 Transcription Factor
  • Trans-Activators
  • RNA Polymerase II