title: Redox potential regulates binding of universal minicircle sequence binding protein at the kinetoplast DNA replication origin.
abstract: Kinetoplast DNA, the mitochondrial DNA of the trypanosomatid Crithidia fasciculata, is a remarkable structure containing 5,000 topologically linked DNA minicircles. Their replication is initiated at two conserved sequences, a dodecamer, known as the universal minicircle sequence (UMS), and a hexamer, which are located at the replication origins of the minicircle L- and H-strands, respectively. A UMS-binding protein (UMSBP), binds specifically the conserved origin sequences in their single stranded conformation. The five CCHC-type zinc knuckle motifs, predicted in UMSBP, fold into zinc-dependent structures capable of binding a single-stranded nucleic acid ligand. Zinc knuckles that are involved in the binding of DNA differ from those mediating protein-protein interactions that lead to the dimerization of UMSBP. Both UMSBP DNA binding and its dimerization are sensitive to redox potential. Oxidation of UMSBP results in the protein dimerization, mediated through its N-terminal domain, with a concomitant inhibition of its DNA-binding activity. UMSBP reduction yields monomers that are active in the binding of DNA through the protein C-terminal region. C. fasciculata trypanothione-dependent tryparedoxin activates the binding of UMSBP to UMS DNA in vitro. The possibility that UMSBP binding at the minicircle replication origin is regulated in vivo by a redox potential-based mechanism is discussed.
authors.affiliation: Department of Parasitology, The Kuvin Center for the Study of Infectious and Tropical Diseases, Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
authors.forename: Itay
authors.initials: I
authors.lastname: Onn
authors.forename: Neta
authors.initials: N
authors.lastname: Milman-Shtepel
authors.forename: Joseph
authors.initials: J
authors.lastname: Shlomai
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D016831:Crithidia fasciculata / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication; D018105:DNA, Kinetoplast / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D019281:Dimerization; D010084:Oxidation-Reduction; D010618:Phenanthrolines / Q000737:chemistry; D011485:Protein Binding; D015800:Protozoan Proteins / Q000378:metabolism*; D018741:Replication Origin; D013879:Thioredoxins / Q000378:metabolism; D015032:Zinc / Q000378:metabolism; D016335:Zinc Fingers
chemical_list: D018105:DNA, Kinetoplast; D004268:DNA-Binding Proteins; D010618:Phenanthrolines; D015800:Protozoan Proteins; C095676:universal minicircle sequence binding protein, protozoan; D013879:Thioredoxins; D015032:Zinc; C025205:1,10-phenanthroline