title: Proteasomal serine hydrolases are up-regulated by and required for influenza virus infection.
abstract: Interactions between viruses and their host cells are important determinants of virus replication and of immune responses to the virus. However, these interactions and resulting consequences of these interactions remain poorly defined. Numerous recent quantitative proteomic approaches have measured host proteins affected by virus infection. Here, we used activity-based protein profiling (ABPP) to measure functional alterations in host serine hydrolases after influenza A virus infection of Madin-Darby canine kidney and human A549 lung cells. We identified 62 serine proteases. We then combined the ABPP approach with stable isotope labeling to directly measure how serine hydrolase activities were affected by virus infection. Differentially regulated SHs mapped into a few key cellular pathway systems, most notably the proteasomal system. The specific serine protease inhibitors Aprotinin and Pefablock and specific proteasomal inhibitors Bortezomib and MG132 significantly inhibited influenza virus growth. Some inhibitors also down-regulated activities of several proteasomal proteins, including PSMA1, PSMA2, and PMSB3. Genetic knockdown of PMSA2 also attenuated influenza virus replication. These findings further our understanding of enzymatic cellular processes affected by influenza virus and may be beneficial in the search for additional antiviral therapeutic targets.
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Manitoba , Winnipeg, Manitoba R3E 0J9, Canada.
authors.forename: Md
authors.initials: M
authors.lastname: Shahiduzzaman
authors.forename: Peyman
authors.initials: P
authors.lastname: Ezatti
authors.forename: Gang
authors.initials: G
authors.lastname: Xin
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Coombs
mesh_terms: D000818:Animals; D007611:Aprotinin / Q000494:pharmacology; D015153:Blotting, Western; D001897:Boronic Acids / Q000494:pharmacology; D000069286:Bortezomib; D045744:Cell Line, Tumor; D004285:Dogs; D054884:Host-Pathogen Interactions / Q000187:drug effects; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D007976:Leupeptins / Q000494:pharmacology; D061985:Madin Darby Canine Kidney Cells; D011480:Protease Inhibitors / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D011719:Pyrazines / Q000494:pharmacology; D034622:RNA Interference; D057057:Serine Proteases / Q000378:metabolism*; D013450:Sulfones / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics
chemical_list: D001897:Boronic Acids; D007976:Leupeptins; D011480:Protease Inhibitors; D020543:Proteome; D011719:Pyrazines; D013450:Sulfones; C002010:4-(2-aminoethyl)benzenesulfonylfluoride; D000069286:Bortezomib; D007611:Aprotinin; D057057:Serine Proteases; D046988:Proteasome Endopeptidase Complex; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde