pr500796n_si_001.pdf (1.78 MB)
Systematic Identification of Hypothetical Bacteriophage Proteins Targeting Key Protein Complexes of Pseudomonas aeruginosa
journal contribution
posted on 2014-10-03, 00:00 authored by 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 LavigneAddressing
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.)