Mutations that inactivate a yeast transcriptional regulatory protein cluster in an evolutionarily conserved DNA binding domain.
The protein encoded by the GAL4 gene of the yeast Saccharomyces cerevisiae binds to DNA upstream of several genes and activates transcription. To try to understand these processes, we have undertaken a genetic analysis of GAL4. Here we report that nearly all missense mutations in GAL4, selected in vivo to lack function of the protein, cluster in the small region of the gene that encodes the DNA binding domain. About half of these mutations alters a cysteine-rich region of the protein highly homologous to several eukaryotic DNA binding proteins; the other half alters some of the 20 amino acids adjacent to the cysteine-rich region. Nearly all of the missense mutations that alter the DNA binding domain abolish the DNA binding activity of GAL4 protein measured in vitro. In contrast, nearly all of the mutations that alter the 3' 95% of the gene that encodes the transcription activation function are nonsense or frameshift mutations. These results support the idea that the conserved cysteine-rich sequence motif is directly involved in binding of several eukaryotic transcriptional regulatory proteins to DNA.
D000595:Amino Acid Sequence; D005075:Biological Evolution; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics*; D005690:Galactose / Q000096:biosynthesis; D005796:Genes; D005800:Genes, Fungal; D005809:Genes, Regulator; D007700:Kinetics; D009154:Mutation; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D005690:Galactose