Survey of Pseudomonas aeruginosa and its phages: de novo peptide sequencing as a novel tool to assess the diversity of worldwide collected viruses.
A collection of 15 newly isolated (bacterio)phages infecting the opportunistic pathogen Pseudomonas aeruginosa was established to investigate their global diversity and potential in phage therapy. These phages were sampled in 14 different countries traversing four continents, from both natural environments and hospital sewage. They all display unique DNA and protein profiles and cluster morphologically into six groups within the three major families of the Caudovirales. Extensive host range studies on a library of 122 AFLP-genotyped clinical P. aeruginosa strains (of which 49 were newly isolated at the University Hospital of Leuven, Belgium) showed that the phages lysed 87% of the strains. Infection analysis of outer membrane mutants identified 10 phages as type IV pili-dependent. More detailed information about the evolutionary relatedness of the phages was gathered by de novo peptide sequencing of major virion proteins using tandem Matrix-Assisted Laser Desorption/Ionization Time of Flight technology. Applying this technique for the first time to viruses, seven groups of closely related phages were identified without the need of prior knowledge of genome content and/or electron microscopic imaging. This study demonstrates both the epidemic population structure of P. aeruginosa and the global spread of P. aeruginosa phage species, and points at the resistance of two clinically predominant, widespread P. aeruginosa strains against phage attack.
Ceyssens
Pieter-Jan
PJ
Division of Gene Technology, Katholieke Universiteit Leuven, Leuven, Belgium.
Noben
Jean-Paul
JP
Ackermann
Hans-W
HW
Verhaegen
Jan
J
De Vos
Daniel
D
Pirnay
Jean-Paul
JP
Merabishvili
Maia
M
Vaneechoutte
Mario
M
Chibeu
Andrew
A
Volckaert
Guido
G
Lavigne
Rob
R
D054458:Amplified Fragment Length Polymorphism Analysis; D015373:Bacterial Typing Techniques; D001530:Belgium; D044822:Biodiversity; D016172:DNA Fingerprinting; D005838:Genotype; D006761:Hospitals; D010449:Peptide Mapping; D011552:Pseudomonas Infections / Q000382:microbiology; D017105:Pseudomonas Phages / Q000737:chemistry* / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification / Q000821:virology*; D020539:Sequence Analysis, Protein / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014771:Virion / Q000648:ultrastructure