A novel DNA-binding motif in the nuclear matrix attachment DNA-binding protein SATB1.
The nuclear matrix attachment DNA (MAR) binding protein SATB1 is a sequence context-specific binding protein that binds in the minor groove, making virtually no contact with the DNA bases. The SATB1 binding sites consist of a special AT-rich sequence context in which one strand is well-mixed A's, T's, and C's, excluding G's (ATC sequences), which is typically found in clusters within different MARs. To determine the extent of conservation of the SATB1 gene among different species, we cloned a mouse homolog of the human STAB1 cDNA from a cDNA expression library of the mouse thymus, the tissue in which this protein is predominantly expressed. This mouse cDNA encodes a 764-amino-acid protein with a 98% homology in amino acid sequence to the human SATB1 originally cloned from testis. To characterize the DNA binding domain of this novel class of protein, we used the mouse SATB1 cDNA and delineated a 150-amino-acid polypeptide as the binding domain. This region confers full DNA binding activity, recognizes the specific sequence context, and makes direct contact with DNA at the same nucleotides as the whole protein. This DNA binding domain contains a novel DNA binding motif: when no more than 21 amino acids at either the N- or C-terminal end of the binding domain are deleted, the majority of the DNA binding activity is lost. The concomitant presence of both terminal sequences is mandatory for binding. These two terminal regions consist of hydrophilic amino acids and share homologous sequences that are different from those of any known DNA binding motifs. We propose that the DNA binding region of SATB1 extends its two terminal regions toward DNA to make direct contact with DNA.
Nakagomi
K
K
La Jolla Cancer Research Foundation, California 92037.
Kohwi
Y
Y
Dickinson
L A
LA
Kohwi-Shigematsu
T
T
D000595:Amino Acid Sequence; D000818:Animals; D034961:Antigens, Nuclear; D001665:Binding Sites; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D003852:Deoxyribonucleoproteins / Q000737:chemistry; D005796:Genes; D036961:Matrix Attachment Region Binding Proteins; D051379:Mice; D008969:Molecular Sequence Data; D015530:Nuclear Matrix / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016415:Sequence Alignment; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D035181:TATA-Box Binding Protein; D013950:Thymus Gland / Q000737:chemistry; D014157:Transcription Factors / Q000737:chemistry
D034961:Antigens, Nuclear; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D003852:Deoxyribonucleoproteins; D036961:Matrix Attachment Region Binding Proteins; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; C076189:SATB1 protein, human; D035181:TATA-Box Binding Protein; D014157:Transcription Factors