The protelomerase of temperate Escherichia coli phage N15 has cleaving-joining activity.
Escherichia coli phage N15 encodes the slightly acidic, 630-residue protein of 72.2 kDa called protelomerase (TelN). TelN is a component of the N15 replication system proposed to be involved in the generation of the linear prophage DNA. This linear DNA molecule has covalently closed ends. The reaction converting circular plasmids into linear molecules was catalyzed in vitro. We demonstrate that the product of telN functions as the protelomerase in the absence of other N15-encoded factors. Purified TelN processes circular and linear plasmid DNA containing the proposed target site telRL to produce linear double-stranded DNA with covalently closed ends. The 56-bp telRL target site consists of a central telO palindrome of 22 bp and two 14-bp flanking sequences comprising inverted repeats. telO is separated from these repeats by 3 bp on each side. The telRL sequence is sufficient for TelN-mediated processing. The ends of the DNA molecules generated in vitro have the same configuration as do those observed in vivo. TelN exerts its activity as cleaving-joining enzyme in a concerted action.
Deneke
J
J
Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, D-14195 Berlin, Germany.
Ziegelin
G
G
Lurz
R
R
Lanka
E
E
D001483:Base Sequence; D003001:Cloning, Molecular; D003090:Coliphages / Q000201:enzymology* / Q000235:genetics; D004261:DNA Replication; D004279:DNA, Viral / Q000378:metabolism*; D004792:Enzyme Precursors / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000821:virology*; D005814:Genes, Viral; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011533:Proviruses; D012091:Repetitive Sequences, Nucleic Acid; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D004279:DNA, Viral; D004792:Enzyme Precursors; D014764:Viral Proteins; C411662:protelomerase protein, Bacteriophage N15; D019098:Telomerase