Binding of transcription factor TFIID to the major late promoter during in vitro nucleosome assembly potentiates subsequent initiation by RNA polymerase II.
A plasmid containing the major late promoter was assembled into nucleosomes in a Xenopus oocyte extract, isolated by gel filtration, and found to be refractory to transcription initiation in vitro. However, exposure of the promoter to HeLa nuclear extract or to a mixture of isolated transcription factors prior to nucleosome assembly prevented nucleosome-mediated repression of the promoter. Inactivation or elimination of the TATA box-binding factor (TFIID) abolished the ability of these treatments to preserve promoter function. Preincubation with TFIID alone prevented repression and resulted in TFIID being sequestered into the nucleosome-assembled templates. Preincubation with all the transcription factors resulted in the assembly of nucleosome templates containing a near complete preinitiation complex, which required only the addition of TFIIE for transcription initiation.
Workman
J L
JL
Laboratory of Biochemistry and Molecular Biology, Rockefeller University, New York, New York 10021.
Roeder
R G
RG
D000818:Animals; D005260:Female; D006367:HeLa Cells / Q000032:analysis; D009707:Nucleosomes / Q000648:ultrastructure*; D009865:Oocytes; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism*; D013698:Templates, Genetic; D014157:Transcription Factors / Q000302:isolation & purification / Q000378:metabolism*; D014158:Transcription, Genetic; D014982:Xenopus laevis
D009707:Nucleosomes; D014157:Transcription Factors; D012319:RNA Polymerase II