title: Interactions of a replication initiator with histone H1-like proteins remodel the condensed mitochondrial genome.
abstract: Kinetoplast DNA (kDNA), the mitochondrial genome of trypanosomatids, consists of several thousand topologically interlocked DNA circles. Mitochondrial histone H1-like proteins were implicated in the condensation of kDNA into a nucleoid structure in the mitochondrial matrix. However, the mechanism that remodels kDNA, promoting its accessibility to the replication machinery, has not yet been described. Analyses, using yeast two hybrid system, co-immunoprecipitation, and protein-protein cross-linking, revealed specific protein-protein interactions between the kDNA replication initiator protein universal minicircle sequence-binding protein (UMSBP) and two mitochondrial histone H1-like proteins. Fluorescence and electron microscopy, as well as biochemical analyses, demonstrated that these protein-protein interactions result in the decondensation of kDNA. UMSBP-mediated decondensation rendered the kDNA network accessible to topological decatenation by topoisomerase II, yielding free kDNA minicircle monomers. Hence, UMSBP has the potential capacity to function in vivo in the activation of the prereplication release of minicircles from the network, a key step in kDNA replication, which precedes and enables its replication initiation. These observations demonstrate the prereplication remodeling of a condensed mitochondrial DNA, which is mediated via specific interactions of histone-like proteins with a replication initiator, rather than through their posttranslational covalent modifications.
authors: Department of Microbiology and Molecular Genetics, The Kuvin Center for the Study of Infectious and Tropical Diseases, Institute of Medical Research Israel-Canada, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
authors: Irit
authors: I
authors: Kapeller
authors: Neta
authors: N
authors: Milman
authors: Nurit
authors: N
authors: Yaffe
authors: Joseph
authors: J
authors: Shlomai
mesh_terms: D016831:Crithidia fasciculata; D004261:DNA Replication; D018105:DNA, Kinetoplast / Q000096:biosynthesis* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D054629:Genome, Mitochondrial / Q000235:genetics*; D006657:Histones / Q000378:metabolism*; D011485:Protein Binding; D015800:Protozoan Proteins / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D018105:DNA, Kinetoplast; D004268:DNA-Binding Proteins; D006657:Histones; C097693:KAP3 protein, Crithidia fasciculata; C097694:KAP4 protein, Crithidia fasciculata; D015800:Protozoan Proteins; C095676:universal minicircle sequence binding protein, protozoan