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PMID: 15987999 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S.

The yeast FACT complex has a role in transcriptional initiation.

Molecular and cellular biology ·Vol. 25 ·No. 14 ·2005-07-00 ·Pages 5812-22

Biswas D, Yu Y, Prall M, Formosa T, Stillman DJ

Abstract

A crucial step in eukaryotic transcriptional initiation is recognition of the promoter TATA by the TATA-binding protein (TBP), which then allows TFIIA and TFIIB to be recruited. However, nucleosomes block the interaction between TBP and DNA. We show that the yeast FACT complex (yFACT) promotes TBP binding to a TATA box in chromatin both in vivo and in vitro. The SPT16 gene encodes a subunit of yFACT, and we show that certain spt16 mutations are synthetically lethal with TBP mutants. Some of these genetic defects can be suppressed by TFIIA overexpression, strongly suggesting a role for yFACT in TBP-TFIIA complex formation in vivo. Mutations in the TOA2 subunit of TFIIA that disrupt TBP-TFIIA complex formation in vitro are also synthetically lethal with spt16. In some cases this spt16 toa2 lethality is suppressed by overexpression of TBP or the Nhp6 architectural transcription factor that is also a component of yFACT. The Spt3 protein in the SAGA complex has been shown to regulate TBP binding at certain promoters, and we show that some spt16 phenotypes can be suppressed by spt3 mutations. Chromatin immunoprecipitations show TBP binding to promoters is reduced in single spt16 and spt3 mutants but increases in the spt16 spt3 double mutant, reflecting the mutual suppression seen in the genetic assays. Finally, in vitro studies show that yFACT promotes TBP binding to a TATA sequence within a reconstituted nucleosome in a TFIIA-dependent manner. Thus, yFACT functions in establishing transcription initiation complexes in addition to the previously described role in elongation.

MeSH Terms
Cell Cycle Proteins/genetics,metabolism,physiology Chromatin Immunoprecipitation Gene Deletion Genes, Lethal Mutation Nucleosomes/metabolism Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins/genetics,metabolism,physiology Suppression, Genetic TATA Box/genetics TATA-Box Binding Protein/metabolism Transcription Factor TFIIA/genetics,metabolism Transcription Factors/genetics,metabolism,physiology Transcription, Genetic/genetics,physiology Transcriptional Elongation Factors
Chemicals
Cell Cycle Proteins Nucleosomes SPT16 protein, S cerevisiae SPT3 protein, S cerevisiae Saccharomyces cerevisiae Proteins TATA-Box Binding Protein TOA2 protein, S cerevisiae Transcription Factor TFIIA Transcription Factors Transcriptional Elongation Factors
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Biswas Debabrata
Department of Pathology, University of Utah Health Sciences Center, 30 North 1900 East, Salt Lake City, Utah 84132-2501, USA.
Yu Yaxin
Prall Matthew
Formosa Tim
Stillman David J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.25.14.5812-5822.2005
Published
2005-07-00
Pages
5812-22
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1168812
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.25.14.5812-5822.2005 PMC Full Text View journal details
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