title: Characterization of newly identified DnaA and DnaB proteins from Acetobacter.
abstract: Although chromosomal replication is an essential feature of the bacterial life cycle, the replication mechanism and involved molecular players have never been properly characterized in the Acetobacter genera. Thanks to whole-genome sequencing, the unknown replication proteins from Acetobacter pasteurianus and Acetobacter orleanensis, DnaA-like and DnaB-like, could be identified. Despite the low nucleotide or amino acid similarity to the respective orthologs from Escherichia coli, their involvement during replication regulation was corroborated by artificial microRNA. In the Acetobacter genome, a novel replication origin, oriAo, was detected with three 9-nucleotide-long DnaA boxes to which DnaA-like proteins bind actively. Bacterial two-hybrid systems and co-immunoprecipitation confirmed the homologous and heterologous interactions between DnaA-like and DnaB-like proteins with their E. coli orthologs. This communication is due to the conserved tryptophan at position 6 for E. coli or 25 for Acetobacter that unables DnaA-like proteins to form oligomeric protein structures after its substitution. Altogether, these results provide novel insights into the genome replication mechanism in Acetobacter.
authors.affiliation: Department of Molecular Biology, Comenius University, Mlynská Dolina B2-122, 842 15 Bratislava, Slovak Republic. Electronic address: bugala@fns.uniba.sk.
authors.forename: Juraj
authors.initials: J
authors.lastname: Bugala
authors.affiliation: Department of Molecular Biology, Comenius University, Mlynská Dolina B2-122, 842 15 Bratislava, Slovak Republic. Electronic address: cimova@fns.uniba.sk.
authors.forename: Viera
authors.initials: V
authors.lastname: Cimová
authors.affiliation: Department of Forest Genetics and Plant Physiology, Umeå Plant Science Center, Swedish University of Agricultural Sciences, 901 83 Umeå, Sweden. Electronic address: peter.grones@slu.se.
authors.forename: Peter
authors.initials: P
authors.lastname: Grones
authors.affiliation: Department of Molecular Biology, Comenius University, Mlynská Dolina B2-122, 842 15 Bratislava, Slovak Republic. Electronic address: grones@fns.uniba.sk.
authors.forename: Jozef
authors.initials: J
authors.lastname: Grones
mesh_terms: D000091:Acetobacter / Q000201:enzymology* / Q000235:genetics; D001665:Binding Sites; D004261:DNA Replication; D004269:DNA, Bacterial / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D047468:Immunoprecipitation; D025941:Protein Interaction Mapping; D018741:Replication Origin; D020798:Two-Hybrid System Techniques
chemical_list: D004269:DNA, Bacterial; D004268:DNA-Binding Proteins
keywords: Acetobacter; DnaA protein; DnaB protein; Protein structure; Protein–protein interaction