Back to Search
Start Over
Site-specific cross-linking of TBP in vivo and in vitro reveals a direct functional interaction with the SAGA subunit Spt3
- Source :
- Genes & Development. 22:2994-3006
- Publication Year :
- 2008
- Publisher :
- Cold Spring Harbor Laboratory, 2008.
-
Abstract
- The TATA-binding protein (TBP) is critical for transcription by all three nuclear RNA polymerases. In order to identify factors that interact with TBP, the nonnatural photoreactive amino acid ρ-benzoyl-phenylalanine (BPA) was substituted onto the surface of Saccharomyces cerevisiae TBP in vivo. Cross-linking of these TBP derivatives in isolated transcription preinitiation complexes or in living cells reveals physical interactions of TBP with transcriptional coregulator subunits and with the general transcription factor TFIIA. Importantly, the results show a direct interaction between TBP and the SAGA coactivator subunits Spt3 and Spt8. Mutations on the Spt3-interacting surface of TBP significantly reduce the interaction of TBP with SAGA, show a corresponding decrease in transcription activation, and fail to recruit TBP to a SAGA-dependent promoter, demonstrating that the direct interaction of these factors is important for activated transcription. These results prove a key prediction of the model for stimulation of transcription at SAGA-dependent genes via Spt3. Our cross-linking data also significantly extend the known surfaces of TBP that directly interact with the transcriptional regulator Mot1 and the general transcription factor TFIIA.
- Subjects :
- Saccharomyces cerevisiae Proteins
Phenylalanine
Protein subunit
genetic processes
Saccharomyces cerevisiae
macromolecular substances
environment and public health
Transcription (biology)
Gene Expression Regulation, Fungal
Coactivator
Genetics
Transcriptional regulation
Promoter Regions, Genetic
Adenosine Triphosphatases
TATA-Binding Protein Associated Factors
biology
General transcription factor
DNA Helicases
TATA-Box Binding Protein
biology.organism_classification
Molecular biology
Cell biology
enzymes and coenzymes (carbohydrates)
Cross-Linking Reagents
Transcription Factor TFIIA
Mutation
Trans-Activators
health occupations
biology.protein
TATA-binding protein
Transcription factor II A
Research Paper
Protein Binding
Transcription Factors
Developmental Biology
Subjects
Details
- ISSN :
- 15495477 and 08909369
- Volume :
- 22
- Database :
- OpenAIRE
- Journal :
- Genes & Development
- Accession number :
- edsair.doi.dedup.....33560ce66bc86d5f472822a62c7006cb
- Full Text :
- https://doi.org/10.1101/gad.1724408