Cloning of a cDNA for DNA Supercoiling Factor Reveals a Distinctive Ca2+-binding Protein

DNA supercoiling factor is a protein capable of generating negative supercoils into a relaxed DNA in conjunction with eukaryotic DNA topoisomerase II. A cDNA clone encoding the Bombyx mori supercoiling factor has been isolated and characterized. The cDNA sequence contains an open reading frame of 322 amino acids rich in acidic residues. A local region of the predicted polypeptide shares a weak homology with the central portion of bacterial DNA gyrase A subunit. The cDNA sequence also predicts two direct repeats within the factor. Each repeat unit consists of a potential EF-hand domain. The factor contains three other potential EF-hand domains and actually binds Ca2+. Moreover, the supercoiling reaction has been shown to be activated by Ca2+. These results implicate Ca2+ in the control of the superhelical state of DNA.