title: Revealing the mode of action of DNA topoisomerase I and its inhibitors by atomic force microscopy.
abstract: In this study, we used, for the first time, atomic force microscope (AFM) images to investigate the mode of action of DNA topoisomerase I (topo I) in the presence and absence of its inhibitors: camptothecin (CPT) and tyrphostin AG-1387. The results revealed that in the absence of the inhibitors, the enzyme relaxed supercoiled DNA starting from a certain point in the DNA molecules and proceeded in one direction towards one of the edges of the DNA molecule. In addition, the relaxation of the supercoiled DNA is subsequently followed by a knotting event. In the presence of CPT, enzyme-supercoiled DNA complexes in which the enzyme is locked inside a relaxed region of the supercoiled DNA molecule were observed. Tyrphostin AG-1387 altered the DNA relaxation process of topo I producing unique shapes of DNA molecules. AFM images of the topo I protein provided a picture of the enzyme, which resembles its known crystallographic structure. Thus, AFM images provide new information on the mode of action of topo I in the absence and presence of its inhibitors.
authors.affiliation: Department of Immunology and Microbiology, Faculty of Health Sciences, The Ben-Gurion Cancer Research Center, Ben-Gurion University, Beer-Sheva, Israel.
authors.forename: Miriam
authors.initials: M
authors.lastname: Argaman
authors.forename: Sarit
authors.initials: S
authors.lastname: Bendetz-Nezer
authors.forename: Sophie
authors.initials: S
authors.lastname: Matlis
authors.forename: Shraga
authors.initials: S
authors.lastname: Segal
authors.forename: Esther
authors.initials: E
authors.lastname: Priel
mesh_terms: D000818:Animals; D002166:Camptothecin / Q000494:pharmacology; D004247:DNA / Q000737:chemistry / Q000648:ultrastructure; D004264:DNA Topoisomerases, Type I / Q000378:metabolism* / Q000648:ultrastructure*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D018625:Microscopy, Atomic Force; D009690:Nucleic Acid Conformation; D020032:Tyrphostins / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D020032:Tyrphostins; D004247:DNA; D004264:DNA Topoisomerases, Type I; D002166:Camptothecin