Molecular genetic dissection of TAF25, an essential yeast gene encoding a subunit shared by TFIID and SAGA multiprotein transcription factors.

Kirchner J, Sanders SL, Klebanow E, Weil PA
Mol Cell Biol. 2001 21 (19): 6668-80

PMID: 11533254 · PMCID: PMC99812 · DOI:10.1128/mcb.21.19.6668-6680.2001

We have performed a systematic structure-function analysis of Saccharomyces cerevisiae TAF25, an evolutionarily conserved, single-copy essential gene which encodes the 206-amino-acid TAF25p protein. TAF25p is an integral subunit of both the 15-subunit general transcription factor TFIID and the multisubunit, chromatin-acetylating transcriptional coactivator SAGA. We used hydroxylamine mutagenesis, targeted deletion, alanine-scanning mutagenesis, high-copy suppression methods, and two-hybrid screening to dissect TAF25. Temperature-sensitive mutant strains generated were used for coimmunoprecipitation and transcription analyses to define the in vivo functions of TAF25p. The results of these analyses show that TAF25p is comprised of multiple mutable elements which contribute importantly to RNA polymerase II-mediated mRNA gene transcription.

MeSH Terms (21)

Amino Acid Sequence Conserved Sequence DNA-Binding Proteins Fungal Proteins Gene Expression Regulation, Fungal Genes, Fungal Macromolecular Substances Molecular Sequence Data Mutation Protein Subunits RNA, Fungal RNA Polymerase II Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Structure-Activity Relationship TATA-Binding Protein Associated Factors Trans-Activators Transcription Factors, TFII Transcription Factor TFIID Two-Hybrid System Techniques

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