Initiation of bacteriophage PRD1 DNA replication on single-stranded templates.
In vitro studies have demonstrated that single-stranded DNA molecules containing the 3' terminal nucleotides of the PRD1 DNA replication origin can support initiation by a protein-primed mechanism. We have determined the minimal requirements for priming by analyzing the template activity of various deletion derivatives. Our results showed that the 3'-terminal 15 nucleotides of the replication origin are sufficient for priming. The finding that the requirements for recognition of replication origin are different from those for priming suggests that there are two distinct steps during initiation of PRD1 DNA replication: first, recognition of the replication origin on double-stranded DNA and second, the priming event on single-stranded DNA. In addition our results showed that additional bases at the 3' end of templates did not affect priming activity, suggesting that the priming site is searched for from inside the template.
Yoo
S K
SK
Department of Microbiology and Immunology, College of Medicine, University of Arizona Tucson 85724.
Ito
J
J
D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D004252:DNA Mutational Analysis; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D013698:Templates, Genetic; D014758:Viral Core Proteins / Q000378:metabolism
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D009841:Oligonucleotides; D014758:Viral Core Proteins