title: The essential role of the 3' terminal template base in the first steps of protein-primed DNA replication.
abstract: Bacteriophages ϕ29 and Nf from Bacillus subtilis start replication of their linear genomes at both ends using a protein-primed mechanism by means of which the DNA polymerase initiates replication by adding dAMP to the terminal protein, this insertion being directed by the second and third 3' terminal thymine of the template strand, respectively. In this work, we have obtained evidences about the role of the 3' terminal base during the initiation steps of ϕ29 and Nf genome replication. The results indicate that the absence of the 3' terminal base modifies the initiation position carried out by ϕ29 DNA polymerase in such a way that now the third position of the template, instead of the second one, guides the incorporation of the initiating nucleotide. In the case of Nf, although the lack of the 3' terminal base has no effect on the initiation position, its absence impairs further elongation of the TP-dAMP initiation product. The results show the essential role of the 3' terminal base in guaranteeing the correct positioning of replication origins at the polymerization active site to allow accurate initiation of replication and further elongation.
authors.affiliation: Instituto de Biología Molecular Eladio Viñuela (Centro Superior de Investigaciones Científicas), Centro de Biología Molecular Severo Ochoa (Centro Superior de Investigaciones Científicas-Universidad Autónoma de Madrid), Cantoblanco, Madrid, Spain.
authors.forename: Irene
authors.initials: I
authors.lastname: Rodríguez
authors.forename: Elisa
authors.initials: E
authors.lastname: Longás
authors.forename: Miguel
authors.initials: M
authors.lastname: de Vega
authors.forename: Margarita
authors.initials: M
authors.lastname: Salas
mesh_terms: D017103:Bacillus Phages / Q000235:genetics; D004261:DNA Replication / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D018741:Replication Origin / Q000235:genetics / Q000502:physiology; D013698:Templates, Genetic
chemical_list: D004259:DNA-Directed DNA Polymerase