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PMID: 11485989 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The S. cerevisiae SAGA complex functions in vivo as a coactivator for transcriptional activation by Gal4.

Genes & development ·Vol. 15 ·No. 15 ·2001-08-01 ·Pages 1946-56

Larschan E, Winston F

Abstract

Previous studies demonstrated that the SAGA (Spt-Ada-Gcn5-Acetyltransferase) complex facilitates the binding of TATA-binding protein (TBP) during transcriptional activation of the GAL1 gene of Saccharomyces cerevisiae. TBP binding was shown to require the SAGA components Spt3 and Spt20/Ada5, but not the SAGA component Gcn5. We have now examined whether SAGA is directly required as a coactivator in vivo by using chromatin immunoprecipitation analysis. Our results demonstrate that SAGA is physically recruited in vivo to the upstream activation sequence (UAS) regions of the galactose-inducible GAL genes. This recruitment is dependent on both induction by galactose and the Gal4 activation domain. Furthermore, we demonstrate that another well-characterized activator, Gal4-VP16, also recruits SAGA in vivo. Finally, we provide evidence that a specific interaction between Spt3 and TBP in vivo is important for Gal4 transcriptional activation at a step after SAGA recruitment. These results, taken together with previous studies, demonstrate a dependent pathway for the recruitment of TBP to GAL gene promoters consisting of the recruitment of SAGA by Gal4 and the subsequent recruitment of TBP by SAGA.

MeSH Terms
Acetyltransferases/metabolism Chromatin/genetics DNA-Binding Proteins/genetics,metabolism Fungal Proteins/genetics,metabolism Gene Expression Regulation, Fungal Genes, Fungal Genotype Histone Acetyltransferases Models, Biological Polymerase Chain Reaction Promoter Regions, Genetic Protein Kinases/metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins TATA-Box Binding Protein Trans-Activators/metabolism Transcription Factors/genetics,metabolism Transcription, Genetic
Chemicals
Chromatin DNA-Binding Proteins Fungal Proteins GAL4 protein, S cerevisiae Saccharomyces cerevisiae Proteins TATA-Box Binding Protein Trans-Activators Transcription Factors Acetyltransferases GCN5 protein, S cerevisiae Histone Acetyltransferases Protein Kinases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Larschan E
Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Winston F
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.911501
Published
2001-08-01
Pages
1946-56
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC312753
Subset
IM
Grants
NIGMS NIH HHS · R01 GM045720 · United States
NIGMS NIH HHS · GM45720 · United States
External Links
PubMed source DOI: 10.1101/gad.911501 PMC Full Text View journal details
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