Abstract
The budding yeast Saccharomyces cerevisiae was used as a model system to study the function of the adenovirus E1A oncoprotein. Previously we demonstrated that expression of the N-terminal 82 amino acids of E1A in yeast causes pronounced growth inhibition and specifically interferes with SWI/SNF-dependent transcriptional activation. Further genetic analysis identified the yeast transcription factor Adr1 as a high copy suppressor of E1A function. Transcriptional activation by Adr1 requires interaction with co-activator proteins Ada2 and Gcn5, components of histone acetyltransferase complexes including ADA and SAGA. Analysis of mutant alleles revealed that several components of the SAGA complex, including proteins from the Ada, Spt, and Taf classes were required for E1A-induced growth inhibition. Growth inhibition also depended on the Gcn5 histone acetyltransferase, and point mutations within the Gcn5 HAT domain rendered cells E1A-resistant. Also required was SAGA component Tra1, a homologue of the mammalian TRRAP protein which is required for c-myc and E1A induced cellular transformation. Additionally, Gcn5 protein could associate with E1A in vitro in a manner that depended on the N-terminal domain of E1A, and Tra1 protein was co-immunoprecipitated with E1A in vivo. These results indicate a strong requirement for intact SAGA complex for E1A to function in yeast, and suggest a role for SAGA-like complexes in mammalian cell transformation.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Acetyltransferases / chemistry
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Acetyltransferases / genetics
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Acetyltransferases / metabolism*
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Adaptor Proteins, Signal Transducing
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Adenovirus E1A Proteins / metabolism*
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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Fungal Proteins / chemistry
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Fungal Proteins / genetics
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Fungal Proteins / metabolism*
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Genes, Fungal / genetics
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Histone Acetyltransferases
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Multienzyme Complexes / chemistry*
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Multienzyme Complexes / genetics
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Multienzyme Complexes / metabolism*
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Protein Binding
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Protein Kinases / chemistry
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Protein Kinases / genetics
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Protein Kinases / metabolism*
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Protein Structure, Tertiary
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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Saccharomyces cerevisiae / enzymology
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism*
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism*
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Suppression, Genetic
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TATA-Binding Protein Associated Factors*
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Trans-Activators / genetics
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Trans-Activators / metabolism
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Transcription Factor TFIID*
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Transfection
Substances
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ADA2 protein, S cerevisiae
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ADR1 protein, S cerevisiae
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Adaptor Proteins, Signal Transducing
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Adenovirus E1A Proteins
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DNA-Binding Proteins
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Fungal Proteins
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HFI1 protein, S cerevisiae
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Multienzyme Complexes
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NGG1 protein, S cerevisiae
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RNA, Messenger
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Saccharomyces cerevisiae Proteins
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TAF5 protein, S cerevisiae
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TATA-Binding Protein Associated Factors
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Trans-Activators
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Transcription Factor TFIID
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Transcription Factors
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Acetyltransferases
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GCN5 protein, S cerevisiae
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Histone Acetyltransferases
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Protein Kinases