DNA-binding properties of the product of the testis-determining gene and a related protein.
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.
Nasrin
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Howard Hughes Medical Institute, Massachusetts General Hospital, Harvard Medical School, Boston 02114.
Buggs
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C
Kong
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XF
Carnazza
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J
Goebl
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Alexander-Bridges
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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*
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