1887

Abstract

Double-stranded DNA phages of both Gram-positive and Gram-negative bacteria typically use a holin–endolysin system to achieve lysis of their host. In this study, the lysis genes of phage P68 were characterized. P68 gene was shown to encode a cell-wall-degrading enzyme, which causes cell lysis when externally added to clinical isolates of . Another gene, , was identified embedded in the −1 reading frame at the 3′ end of . The deduced Hol15 protein has three putative transmembrane domains, and thus resembles class I holins. An additional candidate holin gene, , was found downstream of the endolysin gene based on two predicted transmembrane domains of the encoded protein, which is a typical trait of class II holins. The synthesis of either Hol12 or Hol15 resulted in growth retardation of , and both and were able to complement a phage am mutation. The gene has a dual start motif beginning with the codons Met1-Lys2-Met3…. Evidence is presented that the gene encodes a lysis inhibitor (anti-holin) and a lysis effector (actual holin). As depolarization of the membrane converted the anti-holin to a functional holin, these studies suggested that functions as a typical dual start motif class I holin. The unusual arrangement of the P68 lysis genes is discussed.

Keyword(s): DNP, dinitrophenol
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2005-07-01
2019-11-20
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