Efficient binding of NC2.TATA-binding protein to DNA in the absence of TATA.
Negative cofactor 2 (NC2) forms a stable complex with TATA-binding protein (TBP) on promoters. This prevents the assembly of transcription factor (TF) IIA and TFIIB and leads to repression of RNA polymerase II transcription. Here we have revisited the interactions of NC2.TBP with DNA. We show that NC2.TBP complexes exhibit a significantly reduced preference for TATA box sequences compared with TBP and TBP.TFIIA complexes. In chromatin immunoprecipitations, NC2 is found on a variety of human TATA-containing and TATA-less promoters. Substantial amounts of NC2 are present in a complex with TBP in bulk chromatin. A complex of NC2.TBP displays a K(D) for DNA of approximately 2 x 10(-9) m for a 35-bp major late promoter oligonucleotide. While preferentially recognizing promoter-bound TBP, NC2 also accelerates TBP binding to promoters and stabilizes TBP.DNA complexes. Our data suggest that NC2 controls TBP binding and maintenance on DNA that is largely independent of a canonical TATA sequence.
Gilfillan
Siv
S
Gene Expression, Institute of Molecular Immunology, GSF-National Research Center for Environment and Health, Marchionini-Strasse 25, D-81377 Munich, Germany.
Stelzer
Gertraud
G
Piaia
Elisa
E
Hofmann
Markus G
MG
Meisterernst
Michael
M
D000256:Adenoviridae / Q000235:genetics; D001483:Base Sequence; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D005814:Genes, Viral / Q000235:genetics; D006801:Humans; D007700:Kinetics; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity; D016385:TATA Box / Q000235:genetics*; D035181:TATA-Box Binding Protein / Q000378:metabolism*; D013816:Thermodynamics; D035561:Transcription Factor TFIIA / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*
D002843:Chromatin; D046912:Multiprotein Complexes; D010750:Phosphoproteins; D035181:TATA-Box Binding Protein; D035561:Transcription Factor TFIIA; D014157:Transcription Factors; C075995:down-regulator of transcription 1; D004247:DNA