Studies of bacteriophage P2 DNA replication: localization of the cleavage site of the A protein.
Bacteriophage P2 replicates via a modified rolling circle-type of mechanism, where the P2 A protein acts as an initiator of the replication by inducing a single-stranded cut at the origin of replication (ori). The exact location of the cut induced by the A protein in vivo is determined in this report by: (i) restriction analysis; (ii) DNA sequence analysis; and (iii) primer extensions. It is located 89.2% from the left end of the P2 genome, which is within the coding part of the A gene, in a region devoid of secondary structures. The A gene has been cloned into an expression vector, and the A protein has been purified. The purified A protein does not bind to double-stranded ori containing DNA, but it cleaves single-stranded ori containing DNA, which indicates that a special DNA structure and/or protein is required to make the ori accessible for the A protein.
Liu
Y
Y
Department of Genetics, Stockholm University, Sweden.
Haggård-Ljungquist
E
E
D000595:Amino Acid Sequence; D017901:Bacteriophage P2 / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D003001:Cloning, Molecular; D004261:DNA Replication / Q000502:physiology*; D004277:DNA, Single-Stranded / Q000378:metabolism; D004278:DNA, Superhelical / Q000032:analysis; D004279:DNA, Viral / Q000096:biosynthesis / Q000378:metabolism; D005814:Genes, Viral / Q000235:genetics; D016679:Genome, Viral; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000032:analysis / Q000737:chemistry; D011089:Polydeoxyribonucleotides / Q000378:metabolism; D018741:Replication Origin / Q000235:genetics*; D015183:Restriction Mapping; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000235:genetics; D014779:Virus Replication
D004277:DNA, Single-Stranded; D004278:DNA, Superhelical; D004279:DNA, Viral; D011089:Polydeoxyribonucleotides; D014764:Viral Proteins; D015678:Viral Structural Proteins; C081601:gpA protein, Enterobacteria phage P2