title: Systematic identification of hypothetical bacteriophage proteins targeting key protein complexes of Pseudomonas aeruginosa.
abstract: Addressing the functionality of predicted genes remains an enormous challenge in the postgenomic era. A prime example of genes lacking functional assignments are the poorly conserved, early expressed genes of lytic bacteriophages, whose products are involved in the subversion of the host metabolism. In this study, we focused on the composition of important macromolecular complexes of Pseudomonas aeruginosa involved in transcription, DNA replication, fatty acid biosynthesis, RNA regulation, energy metabolism, and cell division during infection with members of seven distinct clades of lytic phages. Using affinity purifications of these host protein complexes coupled to mass spectrometric analyses, 37 host complex-associated phage proteins could be identified. Importantly, eight of these show an inhibitory effect on bacterial growth upon episomal expression, suggesting that these phage proteins are potentially involved in hijacking the host complexes. Using complementary protein-protein interaction assays, we further mapped the inhibitory interaction of gp12 of phage 14-1 to the α subunit of the RNA polymerase. Together, our data demonstrate the powerful use of interactomics to unravel the biological role of hypothetical phage proteins, which constitute an enormous untapped source of novel antibacterial proteins. (Data are available via ProteomeXchange with identifier PXD001199.).
authors.affiliation: Division of Gene Technology, KU Leuven , B-3001 Leuven, Belgium.
authors.forename: An
authors.initials: A
authors.lastname: Van den Bossche
authors.forename: Pieter-Jan
authors.initials: PJ
authors.lastname: Ceyssens
authors.forename: Jeroen
authors.initials: J
authors.lastname: De Smet
authors.forename: Hanne
authors.initials: H
authors.lastname: Hendrix
authors.forename: Hannelore
authors.initials: H
authors.lastname: Bellon
authors.forename: Nadja
authors.initials: N
authors.lastname: Leimer
authors.forename: Jeroen
authors.initials: J
authors.lastname: Wagemans
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Delattre
authors.forename: William
authors.initials: W
authors.lastname: Cenens
authors.forename: Abram
authors.initials: A
authors.lastname: Aertsen
authors.forename: Bart
authors.initials: B
authors.lastname: Landuyt
authors.forename: Leonid
authors.initials: L
authors.lastname: Minakhin
authors.forename: Konstantin
authors.initials: K
authors.lastname: Severinov
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Noben
authors.forename: Rob
authors.initials: R
authors.lastname: Lavigne
mesh_terms: D000345:Affinity Labels; D001426:Bacterial Proteins / Q000378:metabolism*; D001435:Bacteriophages / Q000378:metabolism*; D015153:Blotting, Western; D002846:Chromatography, Affinity; D011485:Protein Binding; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D000345:Affinity Labels; D001426:Bacterial Proteins; D014764:Viral Proteins
keywords: Bacteriophages; P. aeruginosa; affinity purifications; functional annotation; interactomics