title: Mutational analysis of the DNA-binding domain of yeast heat shock transcription factor.
abstract: Both randomized oligonucleotide cassette mutagenesis and site-directed mutagenesis have been used in combination with a yeast genetic screen to identify critical residues in the DNA-binding domain of heat shock transcription factor from Saccharomyces cerevisiae. Most of the surface residues in this highly conserved domain can be changed to alanine with no observable effect on function. Of nine critical residues identified in this screen, five are within helix alpha 3, previously designated as the probable DNA recognition helix in the crystal structure of the Kluyveromyces lactis protein. The other four residues may be involved in DNA-binding or protein-protein interactions.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley 94720-3206, USA.
authors.forename: S T
authors.initials: ST
authors.lastname: Hubl
authors.forename: J C
authors.initials: JC
authors.lastname: Owens
authors.forename: H C
authors.initials: HC
authors.lastname: Nelson
mesh_terms: D000409:Alanine; D000595:Amino Acid Sequence; D001665:Binding Sites; D017124:Conserved Sequence; D004271:DNA, Fungal / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D006360:Heat-Shock Proteins; D007716:Kluyveromyces / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D004271:DNA, Fungal; D004268:DNA-Binding Proteins; C067168:HSF1 protein, S cerevisiae; D006360:Heat-Shock Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D000409:Alanine