title: Viral terminal protein directs early organization of phage DNA replication at the bacterial nucleoid.
abstract: The mechanism leading to protein-primed DNA replication has been studied extensively in vitro. However, little is known about the in vivo organization of the proteins involved in this fundamental process. Here we show that the terminal proteins (TPs) of phages ϕ29 and PRD1, infecting the distantly related bacteria Bacillus subtilis and Escherichia coli, respectively, associate with the host bacterial nucleoid independently of other viral-encoded proteins. Analyses of phage ϕ29 revealed that the TP N-terminal domain (residues 1-73) possesses sequence-independent DNA-binding capacity and is responsible for its nucleoid association. Importantly, we show that in the absence of the TP N-terminal domain the efficiency of ϕ29 DNA replication is severely affected. Moreover, the TP recruits the phage DNA polymerase to the bacterial nucleoid, and both proteins later are redistributed to enlarged helix-like structures in an MreB cytoskeleton-dependent way. These data disclose a key function for the TP in vivo: organizing the early viral DNA replication machinery at the cell nucleoid.
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), Universidad Autónoma, Canto Blanco, 28049 Madrid, Spain. dmunoz@cbm.uam.es
authors.forename: Daniel
authors.initials: D
authors.lastname: Muñoz-Espín
authors.forename: Isabel
authors.initials: I
authors.lastname: Holguera
authors.forename: David
authors.initials: D
authors.lastname: Ballesteros-Plaza
authors.forename: Rut
authors.initials: R
authors.lastname: Carballido-López
authors.forename: Margarita
authors.initials: M
authors.lastname: Salas
mesh_terms: D017103:Bacillus Phages / Q000235:genetics / Q000502:physiology*; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism / Q000821:virology; D025622:Bacteriophage PRD1 / Q000235:genetics / Q000502:physiology*; D004261:DNA Replication / Q000235:genetics / Q000502:physiology*; D004279:DNA, Viral / Q000096:biosynthesis* / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000821:virology; D005798:Genes, Bacterial; D005814:Genes, Viral; D008954:Models, Biological; D008958:Models, Molecular; D009154:Mutation; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication / Q000235:genetics / Q000502:physiology
chemical_list: C054298:DNA terminal protein, Enterobacteria phage PRD1; D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C040784:terminal protein, Bacillus phage phi29; D004259:DNA-Directed DNA Polymerase
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