Distinct mutations in yeast TAF(II)25 differentially affect the composition of TFIID and SAGA complexes as well as global gene expression patterns.

Kirschner DB, vom Baur E, Thibault C, Sanders SL, Gangloff YG, Davidson I, Weil PA, Tora L
Mol Cell Biol. 2002 22 (9): 3178-93

PMID: 11940675 · PMCID: PMC133751 · DOI:10.1128/MCB.22.9.3178-3193.2002

The RNA polymerase II transcription factor TFIID, composed of the TATA-binding protein (TBP) and TBP-associated factors (TAF(II)s), nucleates preinitiation complex formation at protein-coding gene promoters. SAGA, a second TAF(II)-containing multiprotein complex, is involved in transcription regulation in Saccharomyces cerevisiae. One of the essential protein components common to SAGA and TFIID is yTAF(II)25. We define a minimal evolutionarily conserved 91-amino-acid region of TAF(II)25 containing a histone fold domain that is necessary and sufficient for growth in vivo. Different temperature-sensitive mutations of yTAF(II)25 or chimeras with the human homologue TAF(II)30 arrested cell growth at either the G(1) or G(2)/M cell cycle phase and displayed distinct phenotypic changes and gene expression patterns. Immunoprecipitation studies revealed that TAF(II)25 mutation-dependent gene expression and phenotypic changes correlated at least partially with the integrity of SAGA and TFIID. Genome-wide expression analysis revealed that the five TAF(II)25 temperature-sensitive mutant alleles individually affect the expression of between 18 and 33% of genes, whereas taken together they affect 64% of all class II genes. Thus, different yTAF(II)25 mutations induce distinct phenotypes and affect the regulation of different subsets of genes, demonstrating that no individual TAF(II) mutant allele reflects the full range of its normal functions.

MeSH Terms (23)

Base Composition Blotting, Western Cell Cycle Chromosome Mapping DNA-Binding Proteins Flow Cytometry Fungal Proteins Gene Expression Regulation, Fungal Humans Macromolecular Substances Multiprotein Complexes Mutation Oligonucleotide Array Sequence Analysis Phenotype Recombinant Fusion Proteins Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins TATA-Binding Protein Associated Factors Temperature Transcription Factors Transcription Factors, TFII Transcription Factor TFIID Two-Hybrid System Techniques

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