Identification and characterization of a TFIID-like multiprotein complex from Saccharomyces cerevisiae.
Although the mechanisms of transcriptional regulation by RNA polymerase II are apparently highly conserved from yeast to man, the identification of a yeast TATA-binding protein (TBP)-TBP-associated factor (TAFII) complex comparable to the metazoan TFIID component of the basal transcriptional machinery has remained elusive. Here, we report the isolation of a yeast TBP-TAFII complex which can mediate transcriptional activation by GAL4-VP16 in a highly purified yeast in vitro transcription system. We have cloned and sequenced the genes encoding four of the multiple yeast TAFII proteins comprising the TBP-TAFII multisubunit complex and find that they are similar at the amino acid level to both human and Drosophila TFIID subunits. Using epitope-tagging and immunoprecipitation experiments, we demonstrate that these genes encode bona fide TAF proteins and show that the yeast TBP-TAFII complex is minimally composed of TBP and seven distinct yTAFII proteins ranging in size from M(r) = 150,000 to M(r) = 25,000. In addition, by constructing null alleles of the cloned TAF-encoding genes, we show that normal function of the TAF-encoding genes is essential for yeast cell viability.
Poon
D
D
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232-0615, USA.
Bai
Y
Y
Campbell
A M
AM
Bjorklund
S
S
Kim
Y J
YJ
Zhou
S
S
Kornberg
R D
RD
Weil
P A
PA
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D005800:Genes, Fungal; D006801:Humans; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D035362:Transcription Factor TFIID; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
D035362:Transcription Factor TFIID; D014157:Transcription Factors