Interconversion between active and inactive TATA-binding protein transcription complexes in the mouse genome.
The TATA binding protein (TBP) plays a pivotal role in RNA polymerase II (Pol II) transcription through incorporation into the TFIID and B-TFIID complexes. The role of mammalian B-TFIID composed of TBP and B-TAF1 is poorly understood. Using a complementation system in genetically modified mouse cells where endogenous TBP can be conditionally inactivated and replaced by exogenous mutant TBP coupled to tandem affinity purification and mass spectrometry, we identify two TBP mutations, R188E and K243E, that disrupt the TBP-BTAF1 interaction and B-TFIID complex formation. Transcriptome and ChIP-seq analyses show that loss of B-TFIID does not generally alter gene expression or genomic distribution of TBP, but positively or negatively affects TBP and/or Pol II recruitment to a subset of promoters. We identify promoters where wild-type TBP assembles a partial inactive preinitiation complex comprising B-TFIID, TFIIB and Mediator complex, but lacking TFIID, TFIIE and Pol II. Exchange of B-TFIID in wild-type cells for TFIID in R188E and K243E mutant cells at these primed promoters completes preinitiation complex formation and recruits Pol II to activate their expression. We propose a novel regulatory mechanism involving formation of a partial preinitiation complex comprising B-TFIID that primes the promoter for productive preinitiation complex formation in mammalian cells.
Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, 1 Rue Laurent Fries, 67404 Illkirch Cédex, France.
Mohamed-Amin
MA
Choukrallah
Dominique
D
Kobi
Igor
I
Martianov
W W M Pim
WW
Pijnappel
Nikolai
N
Mischerikow
Tao
T
Ye
Albert J R
AJ
Heck
H Th Marc
HT
Timmers
Irwin
I
Davidson
D000818:Animals; D005786:Gene Expression Regulation; D005816:Genetic Complementation Test; D016678:Genome; D051379:Mice; D009154:Mutation; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D035381:TATA-Binding Protein Associated Factors / Q000737:chemistry / Q000378:metabolism*; D035181:TATA-Box Binding Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D035362:Transcription Factor TFIID / Q000378:metabolism; D014158:Transcription, Genetic; D059467:Transcriptome
D035381:TATA-Binding Protein Associated Factors; D035181:TATA-Box Binding Protein; D035362:Transcription Factor TFIID; D012319:RNA Polymerase II