Dominant negative mutations in yeast TFIID define a bipartite DNA-binding region.
Genetic analysis showed that the conserved C-terminal 180 amino acids of yeast TFIID contain all the essential functions for growth of yeast and response to acidic transcriptional activation signals. A genetic screen was used to identify functionally important residues within this C-terminal region. Five dominant TFIID mutations were isolated that had lost the ability to bind DNA. Four of these mutations were single amino acid substitutions in the most N-terminal of two 66-67 amino acid repeats in TFIID. Analogous mutations made in the most C-terminal repeat all failed to bind DNA and inhibited growth of cells, suggesting that the DNA-binding function of TFIID is partitioned between the two repeated regions. Overproduction of wild-type TFIID rescued the dominance of the TFIID mutants, suggesting that the mutant proteins are dominant because they compete with wild-type TFIID for binding to one or more essential transcription factors.
Reddy
P
P
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Hahn
S
S
D000595:Amino Acid Sequence; D001665:Binding Sites; D015153:Blotting, Western; D004268:DNA-Binding Proteins / Q000235:genetics*; D005799:Genes, Dominant; D005800:Genes, Fungal; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D016385:TATA Box; D035362:Transcription Factor TFIID; D014157:Transcription Factors / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D035362:Transcription Factor TFIID; D014157:Transcription Factors