Human TATA-binding protein-related factor-2 (hTRF2) stably associates with hTFIIA in HeLa cells.
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.
Teichmann
M
M
Laboratory of Biochemistry, The Rockefeller University, New York, NY 10021, USA.
Wang
Z
Z
Martinez
E
E
Tjernberg
A
A
Zhang
D
D
Vollmer
F
F
Chait
B T
BT
Roeder
R G
RG
D000595:Amino Acid Sequence; D000818:Animals; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009842:Oligopeptides; D010455:Peptides; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D012320:RNA Polymerase III / Q000378:metabolism; D012333:RNA, Messenger; D051381:Rats; D035181:TATA-Box Binding Protein; D035341:Telomeric Repeat Binding Protein 2; D013737:Testis / Q000378:metabolism; D035561:Transcription Factor TFIIA; D035362:Transcription Factor TFIID; D014157:Transcription Factors / Q000378:metabolism*; D019473:Transcription Factors, TFII / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation
D004268:DNA-Binding Proteins; D009842:Oligopeptides; D010455:Peptides; D012333:RNA, Messenger; D035181:TATA-Box Binding Protein; D035341:Telomeric Repeat Binding Protein 2; D035561:Transcription Factor TFIIA; D035362:Transcription Factor TFIID; D014157:Transcription Factors; D019473:Transcription Factors, TFII; C085706:FLAG peptide; D012319:RNA Polymerase II; D012320:RNA Polymerase III