Systematic Identification of Hypothetical Bacteriophage Proteins Targeting Key Protein Complexes of Pseudomonas aeruginosa

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....

Full description

Saved in:
Bibliographic Details
Published in:Journal of proteome research 2014-10, Vol.13 (10), p.4446-4456
Main Authors: Van den Bossche, An, Ceyssens, Pieter-Jan, De Smet, Jeroen, Hendrix, Hanne, Bellon, Hannelore, Leimer, Nadja, Wagemans, Jeroen, Delattre, Anne-Sophie, Cenens, William, Aertsen, Abram, Landuyt, Bart, Minakhin, Leonid, Severinov, Konstantin, Noben, Jean-Paul, Lavigne, Rob
Format: Article
Language:English
Subjects:
Affinity Labels
Bacterial Proteins - metabolism
Bacteriophages - metabolism
Blotting, Western
Chromatography, Affinity
Protein Binding
Pseudomonas aeruginosa - metabolism
Tandem Mass Spectrometry
Viral Proteins - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.)
ISSN:1535-3893
1535-3907
DOI:10.1021/pr500796n