Identification and characterization of a TFIID-like multiprotein complex from Saccharomyces cerevisiae

David Poon, Yu Bai, Allyson M. Campbell, Stefan Bjorklund, Young Joon Kim, Sharleen Zhou, Roger D. Kornberg, P. Anthony Weil

Research output: Contribution to journalArticle

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Abstract

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 (TAF(II)) complex comparable to the metazoan TFIID component of the basal transcriptional machinery has remained elusive. Here, we report the isolation of a yeast TBP- TAF(II) 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 TAFn proteins comprising the TBP-TAF(II) 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- TAF(II) complex is minimally composed of TBP and seven distinct yTAF(II) 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.

Original languageEnglish
Pages (from-to)8224-8228
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number18
DOIs
Publication statusPublished - 1995 Aug 29

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Transcription Factor TFIID
TATA-Box Binding Protein
Multiprotein Complexes
Fungal Proteins
Saccharomyces cerevisiae
Yeasts
TATA-Binding Protein Associated Factors
Genes
RNA Polymerase II
Essential Genes
Immunoprecipitation
Protein Binding
Transcriptional Activation
Drosophila
Epitopes
Cell Survival
Proteins
Alleles
Amino Acids

All Science Journal Classification (ASJC) codes

  • Genetics
  • General

Cite this

Poon, David ; Bai, Yu ; Campbell, Allyson M. ; Bjorklund, Stefan ; Kim, Young Joon ; Zhou, Sharleen ; Kornberg, Roger D. ; Weil, P. Anthony. / Identification and characterization of a TFIID-like multiprotein complex from Saccharomyces cerevisiae. In: Proceedings of the National Academy of Sciences of the United States of America. 1995 ; Vol. 92, No. 18. pp. 8224-8228.
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Identification and characterization of a TFIID-like multiprotein complex from Saccharomyces cerevisiae. / Poon, David; Bai, Yu; Campbell, Allyson M.; Bjorklund, Stefan; Kim, Young Joon; Zhou, Sharleen; Kornberg, Roger D.; Weil, P. Anthony.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, No. 18, 29.08.1995, p. 8224-8228.

Research output: Contribution to journalArticle

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