title: Topoisomerase IB of Deinococcus radiodurans resolves guanine quadruplex DNA structures in vitro.
abstract: Deinococcus radiodurans genome contains a large number of guanine repeats interrupted by a few non-guanine bases, termed G motifs. Some of these G motifs were shown forming guanine quadruplex (G4) DNA structure in vitro. How is the formation and relaxation of G4 DNA regulated in the genome of D. radiodurans is not known and is worth investigating. Here, we showed that the topoisomerase Ib of D. radiodurans (DraTopoIB) could change the electrophoretic mobility of fast migrating intramolecular recF-G4 DNA into the slow migrating species. DraTopoIB also reduced the positive ellipticity in circular diachroism (CD) spectra of intramolecular recF-G4 DNA structures stabilized by K+. On the contrary, when DraTopoIB is incubated with G-motifs annealed without K+, it showed neither any change in electrophoretic mobility nor was ellipticity of the CD spectra affected. DNA synthesis by Taq DNA polymerase through G4 DNA structure was attenuated in the presence of G4 DNA binding drugs, which was abrogated by DraTopoIB. This implies that DraTopoIB could destabilize the G4 DNA structure, which is required for G4 drugs binding and stabilization. Camptothecin treatment inhibited DraTopoIB activity on intramolecular G4 DNA structures. These results suggested that DraTopoIB can relax intramolecular G4 DNA structure in vitro and it may be one such protein that could resolve G4 DNA under normal growth conditions in D. radiodurans.
authors.affiliation: Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai 400 084, India.
authors.forename: Swathi
authors.initials: S
authors.lastname: Kota
authors.forename: Hari S
authors.initials: HS
authors.lastname: Misra
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D002166:Camptothecin / Q000494:pharmacology; D002942:Circular Dichroism; D004264:DNA Topoisomerases, Type I / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics; D034301:Deinococcus / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D054856:G-Quadruplexes; D011189:Potassium Chloride / Q000737:chemistry; D059004:Topoisomerase I Inhibitors / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D059004:Topoisomerase I Inhibitors; D011189:Potassium Chloride; C066218:recF protein, Bacteria; D004264:DNA Topoisomerases, Type I; D002166:Camptothecin