Faculty of Pharmaceutical Sciences, University of Tokyo, Bunkyo-ku, Japan.
A DNA-binding protein for the Sarcophaga lectin gene, ATBP (A/T-stretches-binding protein) was purified to homogeneity from the nuclear extract of NIH-Sape-4 cells. The molecular mass of ATBP determined under denaturing conditions was 53 kDa, but its native molecular mass estimated by gel-filtration chromatography was 430 kDa, suggesting that it is an octamer of the 53-kDa subunit. This protein bound to at least three DNA fragments from the Sarcophaga lectin gene, two of them are in the 5'-upstream region and the other is in a region containing an intron. These fragments are very AT rich and inlaid with stretches of A or T residues (A/T stretches). ATBP was found to have affinity for poly[d(A-T)]. These results suggest that ATBP binds to A/T stretches in the three DNA fragments from this gene. Furthermore, longer DNA was found to be bound more effectively, suggesting that an octamer of the 53-kDa subunit has multiple binding sites for the DNA fragments and requires a relatively long DNA sequence for binding.