title: Disclosing the in vivo organization of a viral histone-like protein in Bacillus subtilis mediated by its capacity to recognize the viral genome.
abstract: Organization of replicating prokaryotic genomes requires architectural elements that, similarly to eukaryotic systems, induce topological changes such as DNA supercoiling. Bacteriophage 29 protein p6 has been described as a histone-like protein that compacts the viral genome by forming a nucleoprotein complex and plays a key role in the initiation of protein-primed DNA replication. In this work, we analyze the subcellular localization of protein p6 by immunofluorescence microscopy and show that, at early infection stages, it localizes in a peripheral helix-like configuration. Later, at middle infection stages, protein p6 is recruited to the bacterial nucleoid. This migrating process is shown to depend on the synthesis of components of the 29 DNA replication machinery (i.e., terminal protein and DNA polymerase) needed for the replication of viral DNA, which is required to recruit the bulk of protein p6. Importantly, the double-stranded DNA-binding capacity of protein p6 is essential for its relocalization at the nucleoid. Altogether, the results disclose the in vivo organization of a viral histone-like protein in bacteria.
authors.affiliation: Instituto de Biología Molecular Eladio Viñuela (Consejo Superior de Investigaciones Científicas), Centro de Biología Molecular Severo Ochoa (Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid), Universidad Autónoma, Canto Blanco, 28049 Madrid, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Holguera
authors.forename: David
authors.initials: D
authors.lastname: Ballesteros-Plaza
authors.forename: Daniel
authors.initials: D
authors.lastname: Muñoz-Espín
authors.forename: Margarita
authors.initials: M
authors.lastname: Salas
mesh_terms: D017103:Bacillus Phages / Q000235:genetics* / Q000378:metabolism*; D001412:Bacillus subtilis / Q000166:cytology / Q000821:virology*; D004247:DNA / Q000378:metabolism; D004261:DNA Replication / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics*; D006657:Histones / Q000378:metabolism*; D008954:Models, Biological; D050505:Mutant Proteins / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D006657:Histones; D050505:Mutant Proteins; D014764:Viral Proteins; C413452:phage phi29 protein p6; C040784:terminal protein, Bacillus phage phi29; D004247:DNA