title: Efficient expression and Zn(II)-dependent structure of the DNA binding domain of the yeast GAL4 protein.
abstract: Three protein fragments of different sizes which contain the DNA binding domain of transcription factor GAL4 from Saccharomyces cerevisiae have been expressed in functional forms in Escherichia coli. DNase I footprinting and gel retardation assays showed that the purified proteins bound to the same DNA sequence on the gal1-gal10 promoter as intact GAL4 does. Denaturation--refolding experiments demonstrated that Zn(II) is necessary for maintenance of the conformation of the DNA binding domain of GAL4, as judged on UV-CD and 1H-NMR measurements, as well as for specific DNA binding.
authors.affiliation: Institute for Protein Research, Osaka University, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Serikawa
authors.forename: M
authors.initials: M
authors.lastname: Shirakawa
authors.forename: H
authors.initials: H
authors.lastname: Matsuo
authors.forename: Y
authors.initials: Y
authors.lastname: Kyogoku
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics; D003850:Deoxyribonuclease I; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics*; D015870:Gene Expression; D009682:Magnetic Resonance Spectroscopy; D008667:Metalloproteins / Q000235:genetics; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D014162:Transfection; D015032:Zinc / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D008667:Metalloproteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D004247:DNA; D003850:Deoxyribonuclease I; D015032:Zinc