Sequencing and Characterization of Pseudomonas aeruginosa phage JG004

Abstract

BackgroundPhages could be an important alternative to antibiotics, especially for treatment of multiresistant bacteria as e.g. Pseudomonas aeruginosa. For an effective use of bacteriophages as antimicrobial agents, it is important to understand phage biology but also genes of the bacterial host essential for phage infection.ResultsWe isolated and characterized a lytic Pseudomonas aeruginosa phage, named JG004, and sequenced its genome. Phage JG004 is a lipopolysaccharide specific broad-host-range phage of the Myoviridae phage family. The genome of phage JG004 encodes twelve tRNAs and is highly related to the PAK-P1 phage genome. To investigate phage biology and phage-host interactions, we used transposon mutagenesis of the P. aeruginosa host and identified P. aeruginosa genes, which are essential for phage infection. Analysis of the respective P. aeruginosa mutants revealed several characteristics, such as host receptor and possible spermidine-dependance of phage JG004.ConclusionsWhole genome sequencing of phage JG004 in combination with identification of P. aeruginosa host genes essential for infection, allowed insights into JG004 biology, revealed possible resistance mechanisms of the host bacterium such as mutations in LPS and spermidine biosynthesis and can also be used to characterize unknown gene products in P. aeruginosa.

Adaptive Divergence in Experimental Populations ofPseudomonas fluorescens. V. Insight into the Niche Specialist Fuzzy Spreader Compels Revision of the ModelPseudomonasRadiation

“…Indeed, O antigen is essential for the establishment of infection (or protection against host defenses) in a diverse array of plant and animal pathogens (Cava et al 1989;Dekkers et al 1998;Yang et al 2000;Kannenberg and Carlson 2001;Poon et al 2008;Kesawat et al 2009); moreover, the O antigen serves as a receptor for phages (Michel et al 2010;Filippov et al 2011;Garbe et al 2011;Shin et al 2012). Thus it would appear that O antigens are subject to two conflicting selective pressures: pressure to change to avoid phage predation and pressure to be maintained to enable colonization of host species during infection.…”

“…Single deletions of genes fuzVWX and fuzZ (but not fuzX) also conferred resistance to SBW25F2 (Table 3). Rough fuzW mutants (PBR746-R) are phage resistant whereas smooth fuzW mutants (PBR746-S) are phage sensitive.Location of the FS causal mutation to a gene product predicted to modify LPS allows us to speculate upon the F2 receptor: LPS serves as a receptor for a broad range of bacteriophage in many bacterial species (for examples see Michel et al 2010;Filippov et al 2011;Garbe et al 2011;Shin et al 2012). It appears likely that phage resistance in FS is conferred by alterations in LPS structure.…”

High Diversity and Novel Species of Pseudomonas aeruginosa Bacteriophages

“…S4 in the supplemental material). Twenty of them were compiled previously (12); another two species included the characterized phages, KPP1, JG004, and PAK_P1 (18,20), and the other six reported here (PaMx11, PaMx25, PaMx28, PaMx31, PaMx42, and PaMx74) were new because they did not have homologues in GenBank.…”

A Novel Cyanophage with a Cyanobacterial Nonbleaching Protein A Gene in the Genome

“…For examples, some photosynthesis-related genes (psbA, hliP, and PSII) and stress-response genes (coding for chaperones and genes associated with bacterial motility and chemotaxis) have been described in cyanophages (4, 5, 31, 36, 47), most of which are transcribed together with essential cyanophage replication-related genes (6, 13, 30, 65). Moreover, a nonbleaching protein A (NblA) gene has been found from a lytic phage, Ma-LMM01, infecting Microcystis aeruginosa (50, 64), but its function has not been demonstrated.Despite different hosts and habitats, more than 95% of the known cyanophages belong to the tailed phages (16,68). The tailed phages are classified into three families: Myoviridae with long contractile tails, Siphoviridae with long noncontractile tails, and Podoviridae with short tails.…”

“…Despite different hosts and habitats, more than 95% of the known cyanophages belong to the tailed phages (16,68). The tailed phages are classified into three families: Myoviridae with long contractile tails, Siphoviridae with long noncontractile tails, and Podoviridae with short tails.…”