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12. |
- Poon, D, et al.
(author)
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Identification and characterization of a TFIID-like multiprotein complex from Saccharomyces cerevisiae.
- 1995
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In: Proceedings of the National Academy of Sciences of the United States of America. - 0027-8424 .- 1091-6490. ; 92:18, s. 8224-8
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Journal article (peer-reviewed)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 (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.
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13. |
- Björklund, Stefan, et al.
(author)
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Mediator of transcriptional regulation.
- 1996
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In: TIBS -Trends in Biochemical Sciences. Regular ed.. - 0968-0004 .- 1362-4326. ; 21:9, s. 335-7
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Journal article (peer-reviewed)abstract
- A multi-protein complex, termed mediator, has been isolated from yeast, based on its requirement for transcriptional activation in a system reconstituted from pure RNA polymerase II and general transcription factors. Mediator polypeptides include the products of many genes previously recovered from screens for mutations affecting transcription. This connection between biochemical and genetic studies reveals that mediator is important for both activation and repression of transcription, and that mediator plays a role in transcriptional regulation in vivo as well as in vitro. Mediator binds the carboxy-terminal domain of RNA polymerase II, forming a polymerase holoenzyme, whose possible association with additional proteins is a subject of some controversy.
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14. |
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