title: Genome analysis of the staphylococcal temperate phage DW2 and functional studies on the endolysin and tail hydrolase.
abstract: This study describes the genome of temperateSiphoviridaephage DW2, which is routinely propagated onStaphylococcus aureusDPC5246. The 41941 bp genome revealed an open reading frame (ORF1) which has a high level of homology with members of the resolvase subfamily of site-specific serine recombinase, involved in chromosomal integration and excision. In contrast, the majority of staphylococcal phages reported to date encode tyrosine recombinases. Two putative genes encoded by phage DW2 (ORF15 and ORF24) were highly homologous to the NWMN0273 and NWMN0280 genes encoding virulence factors carried on the genome of ϕNM4, a prophage in the genome ofS. aureusNewman. Phage DW2 also encodes proteins highly homologous to two well-characterizedStaphylococcus aureuspathogenicity island derepressors encoded by the staphylococcal helper phage 80α indicating that it may similarly act as a helper phage for mobility of pathogenicity islands inS. aureus. This study also focused on the enzybiotic potential of phage DW2. The structure of the putative endolysin and tail hydrolase were investigated and used as the basis for a cloning strategy to create recombinant peptidoglycan hydrolyzing proteins. After overexpression inE. coli,four of these proteins (LysDW2, THDW2, CHAPE1-153, and CHAPE1-163) were demonstrated to have hydrolytic activity against peptidoglycan ofS. aureusand thus represent novel candidates for exploitation as enzybiotics.
authors.affiliation: Department of Biological Sciences; Cork Institute of Technology; Bishopstown, Cork Ireland.
authors.forename: Ruth
authors.initials: R
authors.lastname: Keary
authors.affiliation: Biotechnology Department; Teagasc; Moorepark Food Research Centre; Fermoy, Co. Cork Ireland.
authors.forename: Olivia
authors.initials: O
authors.lastname: McAuliffe
authors.affiliation: Biotechnology Department; Teagasc; Moorepark Food Research Centre; Fermoy, Co. Cork Ireland.
authors.forename: R Paul
authors.initials: RP
authors.lastname: Ross
authors.affiliation: Alimentary Pharmacobiotic Centre; University College Cork; Cork, Ireland.
authors.forename: Colin
authors.initials: C
authors.lastname: Hill
authors.affiliation: Department of Biological Sciences; Cork Institute of Technology; Bishopstown, Cork Ireland.
authors.forename: Jim
authors.initials: J
authors.lastname: O'Mahony
authors.affiliation: Department of Biological Sciences; Cork Institute of Technology; Bishopstown, Cork Ireland.
authors.forename: Aidan
authors.initials: A
authors.lastname: Coffey
keywords: Staphylococcus; bacteriophage; endolysin; serine recombinase; virion-associated peptidoglycan hydrolase; virulence
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