Molecular and physiological analysis of three Pseudomonas aeruginosa phages belonging to the “N4-like viruses”

Abstract

We present a detailed analysis of the genome architecture, structural proteome and infection-related properties of three Pseudomonas phages, designated LUZ7, LIT1 and PEV2. These podoviruses encapsulate 72.5 to 74.9 kb genomes and lyse their host after 25 min aerobic infection. PEV2 can successfully infect under anaerobic conditions, but its latent period is tripled, the lysis proceeds far slower and the burst size decreases significantly. While the overall genome structure of these phages resembles the well-studied coliphage N4, these Pseudomonas phages encode a cluster of tail genes which displays significant similarity to a Pseudomonasaeruginosa (cryptic) prophage region. Using ESI-MS/MS, these tail proteins were shown to be part of the phage particle, as well as ten other proteins including a giant 370 kDa virion RNA polymerase. These phages are the first described representatives of a novel kind of obligatory lytic P. aeruginosa-infecting phages, belonging to the widespread “N4-like viruses” genus.

Contrasted coevolutionary dynamics between a bacterial pathogen and its bacteriophages

“…10 7 cfu/mL at −80°C. Of the six bacteriophage strains used (Table S1), four (LKD16, PEV2, LUZ19, and LUZ7) were from the Podoviridae, and two (14/1 and LMA2) were from the Myoviridae (64)(65)(66). LUZ7 and PEV2 have 70-nm icosahedral capsids and short tails, typical of the Podoviridae.…”

“…LUZ7 and PEV2 have 70-nm icosahedral capsids and short tails, typical of the Podoviridae. PEV2 is capable of infecting PA01 both aerobically and anaerobically, an important consideration for clinical applications in which infections can be anaerobic in thick mucus (65). LUZ19 and LKD16 are slightly smaller, with head diameters of 65 and 60 nm, respectively.…”

Development of Giant Bacteriophage ϕKZ Is Independent of the Host Transcription Apparatus

“…The N4-like viruses have evolved an inverse transcriptional strategy. An ssRNAP is carried within their capsids and is injected with phage DNA upon infection to transcribe early phage promoters (17)(18)(19). A second N4-encoded ssRNAP, a product of an early gene, transcribes middle phage genes.…”

In vitro design of a novel lytic bacteriophage cocktail with therapeutic potential against organisms causing diabetic foot infections

“…EU418428) and GH15 (Gu et al, 2012). The bacteriophages F510/08 and F770/05 shared high sequence identity (up to 98 % nucleotide sequence identity, 83-98 % genome coverage) with Pseudomonas wKMV-like and N4-like viruses, respectively (Ceyssens et al, 2010). Examples of wKMV-like viruses are the bacteriophages wKMV and LUZ19 (Ceyssens et al, 2006;Lavigne et al, 2003) and of N4-like viruses are LIT1 and LUZ7 (Ceyssens et al, 2010).…”

“…The bacteriophages F510/08 and F770/05 shared high sequence identity (up to 98 % nucleotide sequence identity, 83-98 % genome coverage) with Pseudomonas wKMV-like and N4-like viruses, respectively (Ceyssens et al, 2010). Examples of wKMV-like viruses are the bacteriophages wKMV and LUZ19 (Ceyssens et al, 2006;Lavigne et al, 2003) and of N4-like viruses are LIT1 and LUZ7 (Ceyssens et al, 2010). The bacteriophage F1245/05 presented no significant similarity at the DNA level with any known bacteriophage in the databases, except for a few short segments with up to 4 % nucleotide sequence identity and 81 % genome coverage.…”