In vitro replication of bacteriophage PRD1 DNA. Metal activation of protein-primed initiation and DNA elongation.
Bacteriophage PRD1 replicates its DNA by means of a protein-primed replication mechanism. Compared to Mg2+, the use of Mn2+ as the metal activator of the phage DNA polymerase results in a great stimulation of the initiation reaction. The molecular basis of the observed stimulatory effect is an increase in the velocity of the reaction. The phage DNA polymerase is also able to catalyze the formation of the initiation complex in the absence of DNA template. Although the presence of Mn2+ does not affect either the polymerization activity or the processivity of the DNA polymerase, this metal is unable to activate the overall replication of the phage genome. This can be explained by a deleterious effect of Mn2+ on the 3'-5'-exonucleolytic and/or the strand-displacement activity, the latter being an intrinsic function of the viral DNA polymerase required for protein-primed DNA replication.
Caldentey
J
J
Department of Genetics, University of Helsinki, Finland.
Blanco
L
L
Savilahti
H
H
Bamford
D H
DH
Salas
M
M
D001435:Bacteriophages / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D002412:Cations / Q000378:metabolism; D004261:DNA Replication / Q000502:physiology*; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D005092:Exonucleases / Q000378:metabolism; D008670:Metals / Q000378:metabolism*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
D002412:Cations; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D008670:Metals; D009838:Oligodeoxyribonucleotides; D014764:Viral Proteins; D004259:DNA-Directed DNA Polymerase; D005092:Exonucleases