title: Phage T4-induced DNA breaks activate a tRNA repair-defying anticodon nuclease.
abstract: The natural role of the conserved bacterial anticodon nuclease (ACNase) RloC is not known, but traits that set it apart from the homologous phage T4-excluding ACNase PrrC could provide relevant clues. PrrC is silenced by a genetically linked DNA restriction-modification (RM) protein and turned on by a phage-encoded DNA restriction inhibitor. In contrast, RloC is rarely linked to an RM protein, and its ACNase is regulated by an internal switch responsive to double-stranded DNA breaks. Moreover, PrrC nicks the tRNA substrate, whereas RloC excises the wobble nucleotide. These distinctions suggested that (i) T4 and related phage that degrade their host DNA will activate RloC and (ii) the tRNA species consequently disrupted will not be restored by phage tRNA repair enzymes that counteract PrrC. Consistent with these predictions we show that Acinetobacter baylyi RloC expressed in Escherichia coli is activated by wild-type phage T4 but not by a mutant impaired in host DNA degradation. Moreover, host and T4 tRNA species disrupted by the activated ACNase were not restored by T4's tRNA repair system. Nonetheless, T4's plating efficiency was inefficiently impaired by AbaRloC, presumably due to a decoy function of the phage encoded tRNA target, the absence of which exacerbated the restriction.
authors.affiliation: Department of Biochemistry and Molecular Biology, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Lital
authors.initials: L
authors.lastname: Bitton
authors.affiliation: Department of Biochemistry and Molecular Biology, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Daniel
authors.initials: D
authors.lastname: Klaiman
authors.affiliation: Department of Biochemistry and Molecular Biology, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Kaufmann
mesh_terms: D000150:Acinetobacter / Q000235:genetics / Q000821:virology; D000595:Amino Acid Sequence; D017122:Bacteriophage T4 / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity; D053960:DNA Breaks; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics / Q000821:virology; D029968:Escherichia coli Proteins / Q000235:genetics; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism*; D012260:Ribonucleases / Q000235:genetics / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; D012343:RNA, Transfer; C084965:PrrC protein, E coli; D012260:Ribonucleases; C049345:anticodon nuclease