Cloning and expression of human TAFII250: a TBP-associated factor implicated in cell-cycle regulation.
Basal transcription by human RNA polymerase II requires the coordinate action of several ancillary factors (TFIIA-J) and can be regulated by various promoter-specific DNA binding proteins. An additional class of factors, called coactivators, are dispensable for basal transcription but are indispensable for regulation by transcriptional activators. Biochemical studies established that some coactivators are associated with the TATA-binding protein (TBP) to form the TFIID complex. We therefore set out to define the relationship between TBP and these TBP-associated factors (TAFs). Here we describe the cloning, expression and properties of the first human TAF, hTAFII250. The hTAFII250 gene is identical to a gene, CCG1, (ref 7,8), implicated in cell-cycle progression. Recombinant hTAFII250 binds directly to TBP both in vitro and in yeast, and participates in the formation of the TFIID complex. This largest TAF may therefore play a central role in TFIID assembly by interacting with both TBP and other TAFs, as well as serving to link the control of transcription to the cell cycle.
Ruppert
S
S
Howard Hughes Medical Institute, Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Wang
E H
EH
Tjian
R
R
D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle / Q000502:physiology*; D002460:Cell Line; D002846:Chromatography, Affinity; D003001:Cloning, Molecular / Q000379:methods; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D006367:HeLa Cells; D051548:Histone Acetyltransferases; D006801:Humans; D007313:Insecta; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D009838:Oligodeoxyribonucleotides; D011061:Poly A / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger; D011994:Recombinant Proteins / Q000378:metabolism; D016385:TATA Box; D035381:TATA-Binding Protein Associated Factors; D035181:TATA-Box Binding Protein; D035362:Transcription Factor TFIID; D014157:Transcription Factors / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D014162:Transfection
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D011994:Recombinant Proteins; D035381:TATA-Binding Protein Associated Factors; D035181:TATA-Box Binding Protein; D035362:Transcription Factor TFIID; D014157:Transcription Factors; D011061:Poly A; D012313:RNA; D051548:Histone Acetyltransferases; C069001:TATA-binding protein associated factor 250 kDa