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: An Van den Bossche, Pieter-Jan Ceyssens, Jeroen De Smet, Hanne Hendrix, Hannelore Bellon, Nadja Leimer, Jeroen Wagemans, Anne-Sophie Delattre, William Cenens, Abram Aertsen, Bart Landuyt, Leonid Minakhin, Konstantin Severinov, Jean-Paul Noben, Rob Lavigne
• Journal: Journal of proteome research (J Proteome Res) Vol. 13 Issue 10 Pg. 4446-56 (Oct 03 2014) ISSN: 1535-3907 [Electronic] United States
• PMID: 25185497 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Affinity Labels
• Bacterial Proteins
• Viral Proteins

Topics

• Affinity Labels
• Bacterial Proteins (metabolism)
• Bacteriophages (metabolism)
• Blotting, Western
• Chromatography, Affinity
• Protein Binding
• Pseudomonas aeruginosa (metabolism)
• Tandem Mass Spectrometry
• Viral Proteins (metabolism)