The BRCA1 and BARD1 association with the RNA polymerase II holoenzyme

Cancer Res. 2002 Aug 1;62(15):4222-8.

Abstract

We have previously shown that endogenous BRCA1 and overexpressed epitope-tagged BRCA1 are present in the transcription complex called the RNA polymerase II holoenzyme (holo-pol). In this study, we further characterized BRCA1 association with the holo-pol by overexpressing deletion mutants of epitope-tagged BRCA1. We found that BRCA1-associated RING domain protein (BARD1) is a component of the holo-pol complex. Deletion of the BRCA1 NH(2) terminus, which is bound by BARD1 as well as other proteins, eliminates >98% of BRCA1 association with the holo-pol. In contrast with earlier observations, deletion of the COOH terminus of BRCA1 did not affect significantly the association with holo-pol. Immunocytochemistry of expressed full-length and deletion mutants of BRCA1 showed that the NH(2) terminus of BRCA1 is important for nuclear dot formation in S-phase. An intact BRCA1 NH(2) terminus is required for the association with holo-pol and for subnuclear localization in S-phase foci. Taken together, these data support a role for BRCA1 regulation of holo-pol function.

Publication types

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

MeSH terms

  • Adenoviridae / genetics
  • BRCA1 Protein / genetics
  • BRCA1 Protein / metabolism*
  • Carrier Proteins / metabolism*
  • Cell Line
  • Epitopes
  • Gene Deletion
  • Holoenzymes / metabolism
  • Humans
  • Immunohistochemistry
  • RNA Polymerase II / metabolism*
  • S Phase / physiology
  • Tumor Suppressor Proteins*
  • Ubiquitin-Protein Ligases*

Substances

  • BRCA1 Protein
  • Carrier Proteins
  • Epitopes
  • Holoenzymes
  • Tumor Suppressor Proteins
  • BARD1 protein, human
  • Ubiquitin-Protein Ligases
  • RNA Polymerase II