title: DNA-binding properties of the product of the testis-determining gene and a related protein.
abstract: THE upstream region of the human glyceraldehyde-3-phosphate dehydrogenase gene contains an insulin-response element (IRE-A) responsible for insulin-dependent transcription of the gene. The open reading frame of a rat complementary DNA encoding a protein (IRE-ABP) that binds to this sequence contains an HMG box motif that is 67% identical to the mouse candidate gene for the testis-determining factor SRY, and 98% identical to the mouse SRY-like gene, a4. Here we report that IRE-ABP and SRY bind to IRE-A DNA with comparable specificity in a DNase-I footprinting assay. Two females with sex reversal were found to have a single amino-acid substitution in the HMG box domain of SRY at position 3 and 7, respectively. SRY derivatives containing corresponding mutations do not make contact with IRE-A DNA. These results are direct evidence that mouse SRY-like proteins are sequence-specific DNA-binding proteins and identify two amino acids critical to this interaction. Moreover, IRE-A is a candidate SRY-response element.
authors.affiliation: Howard Hughes Medical Institute, Massachusetts General Hospital, Harvard Medical School, Boston 02114.
authors.forename: N
authors.initials: N
authors.lastname: Nasrin
authors.forename: C
authors.initials: C
authors.lastname: Buggs
authors.forename: X F
authors.initials: XF
authors.lastname: Kong
authors.forename: J
authors.initials: J
authors.lastname: Carnazza
authors.forename: M
authors.initials: M
authors.lastname: Goebl
authors.forename: M
authors.initials: M
authors.lastname: Alexander-Bridges
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000502:physiology*; D005786:Gene Expression Regulation; D006801:Humans; D007328:Insulin / Q000502:physiology; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D009841:Oligonucleotides / Q000737:chemistry; D011994:Recombinant Proteins; D012045:Regulatory Sequences, Nucleic Acid; D051876:Sex-Determining Region Y Protein; D014157:Transcription Factors / Q000502:physiology*
chemical_list: D004268:DNA-Binding Proteins; D007328:Insulin; D009687:Nuclear Proteins; D009841:Oligonucleotides; D011994:Recombinant Proteins; C497745:SRY protein, human; D051876:Sex-Determining Region Y Protein; D014157:Transcription Factors