title: Characterization of recombinant human DNA topoisomerase IIIalpha activity expressed in yeast.
abstract: Recombinant human DNA topoisomerase IIIalpha was expressed in mutant yeast cells devoid of both topoisomerases I and III, and the gene product was partially purified. The activity of the protein in supercoil removal was found to be limited and also biphasic: in the first phase it processively changed the linking-number of hypernegatively supercoiled DNA, but only to the superhelicity of a regular negative supercoil; in the second phase the enzyme relaxed the DNA further, but only slightly and slowly. The optimal solution conditions for the enzyme activity were found to be physiological. The assay results with a truncation mutant showed that the C-terminal 334 amino acids are unnecessary for the activity, suggesting that this region, and perhaps the entire protein, is involved in a function other than supercoil removal.
authors.affiliation: Department of Chemistry, College of Science, Rikkyo (St. Paul's) University, Nishi-Ikebukuro, Toshima-ku, Tokyo 171-8501, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Hotoda
authors.forename: R
authors.initials: R
authors.lastname: Hanai
mesh_terms: D004264:DNA Topoisomerases, Type I / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D004278:DNA, Superhelical / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D009154:Mutation; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics
chemical_list: D004278:DNA, Superhelical; D011994:Recombinant Proteins; D004264:DNA Topoisomerases, Type I