Open AccessFunctional elucidation of antibacterial phage ORFans targeting P seudomonas aeruginosa
Author(s) -
Wagemans Jeroen,
Blasdel Bob G.,
Van den Bossche An,
Uytterhoeven Birgit,
De Smet Jeroen,
Paeshuyse Jan,
Cenens William,
Aertsen Abram,
Uetz Peter,
Delattre AnneSophie,
Ceyssens PieterJan,
Lavigne Rob
Publication year - 2014
Publication title -
cellular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.542
H-Index - 138
eISSN - 1462-5822
pISSN - 1462-5814
DOI - 10.1111/cmi.12330
Subject(s) - biology , bacteriophage , pseudomonas aeruginosa , microbiology and biotechnology , spermidine , virulence , phagemid , phage display , type vi secretion system , phage therapy , genome , bacteria , dna , gene , genetics , escherichia coli , enzyme , biochemistry , antibody
Summary Immediately after infection, virulent bacteriophages hijack the molecular machinery of their bacterial host to create an optimal climate for phage propagation. For the vast majority of known phages, it is completely unknown which bacterial functions are inhibited or coopted. Early expressed phage genome regions are rarely identified, and often filled with small genes with no homology in databases (so‐called ORFans ). In this work, we first analysed the temporal transcription pattern of the N4 ‐like Pseudomonas ‐infecting phages and selected 26 unknown, early phage ORFans . By expressing their encoded proteins individually in the host bacterium P seudomonas aeruginosa , we identified and further characterized six antibacterial early phage proteins using time‐lapse microscopy, radioactive labelling and pull‐down experiments. Yeast two‐hybrid analysis gaveclues to their possible role in phage infection. Specifically, we show that the inhibitory proteins may interact with transcriptional regulator PA 0120, the replicative DNA helicase DnaB , the riboflavin metabolism key enzyme RibB , the ATPase PA0657 and the spermidine acetyltransferase PA4114 . The dependency of phage infection on spermidine was shown in a final experiment. In the future, knowledge of how phages shut down their hosts as well ass novel phage–host interaction partners could be very valuable in the identification of novel antibacterial targets.