Human TATA-binding protein-related factor-2 (hTRF2) stably associates with hTFIIA in HeLa cells

Proc Natl Acad Sci U S A. 1999 Nov 23;96(24):13720-5. doi: 10.1073/pnas.96.24.13720.

Abstract

The TATA-binding protein (TBP)-related factor TRF1, has been described in Drosophila and a related protein, TRF2, has been found in a variety of higher eukaryotes. We report that human (h)TRF2 is encoded by two mRNAs with common protein coding but distinct 5' nontranslated regions. One mRNA is expressed ubiquitously (hTRF2-mRNA1), whereas the other (hTRF2-mRNA2) shows a restricted expression pattern and is extremely abundant in testis. In addition, we show that hTRF2 forms a stable stoichiometric complex with hTFIIA, but not with TAFs, in HeLa cells stably transfected with flag-tagged hTRF2. Neither recombinant human (rh)TRF2 nor the native flag.hTRF2-TFIIA complex is able to replace TBP or TFIID in basal or activated transcription from various RNA polymerase II promoters. Instead, rhTRF2, but not the flag.hTRF2-TFIIA complex, moderately inhibits basal or activated transcription in the presence of rhTBP or flag.TFIID. This effect is either completely (TBP-mediated transcription) or partially (TFIID-mediated transcription) counteracted by addition of free TFIIA. Neither rhTRF2 nor flag. hTRF2-TFIIA has any effect on the repression of TFIID-mediated transcription by negative cofactor-2 (NC2) and neither substitutes for TBP in RNA polymerase III-mediated transcription.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • DNA-Binding Proteins* / genetics
  • DNA-Binding Proteins* / metabolism*
  • HeLa Cells
  • Humans
  • Male
  • Molecular Sequence Data
  • Oligopeptides
  • Peptides
  • Promoter Regions, Genetic
  • RNA Polymerase II / metabolism
  • RNA Polymerase III / metabolism
  • RNA, Messenger
  • Rats
  • TATA-Box Binding Protein
  • Telomeric Repeat Binding Protein 2
  • Testis / metabolism
  • Transcription Factor TFIIA
  • Transcription Factor TFIID
  • Transcription Factors* / metabolism*
  • Transcription Factors, TFII / metabolism
  • Transcription, Genetic
  • Transcriptional Activation

Substances

  • DNA-Binding Proteins
  • Oligopeptides
  • Peptides
  • RNA, Messenger
  • TATA-Box Binding Protein
  • Telomeric Repeat Binding Protein 2
  • Transcription Factor TFIIA
  • Transcription Factor TFIID
  • Transcription Factors
  • Transcription Factors, TFII
  • FLAG peptide
  • RNA Polymerase II
  • RNA Polymerase III