title: Suppression of apoptosis by pseudorabies virus Us3 protein kinase through the activation of PI3-K/Akt and NF-κB pathways.
abstract: The pseudorabies virus (PRV) is a major viral disease that causes huge economic loss in the pig industry globally. Most viruses have been found to generate anti-apoptotic factors that facilitate cell survival in the early stages of infection. This study aimed to investigate the anti-apoptotic effects of PRV and study the underlying mechanisms in the early stage of infection. We investigated and compared whether the two PRV Us3 isoforms, Us3a and Us3b, could block apoptosis induced by virus infection, and further identified molecules involved in the signaling pathways. Our results demonstrated that PRV elicits 3-phosphoinositide dependent protein kinase-1/phosphatidylinositide 3-kinases/Akt (PDK-1/PI3-K/Akt)- and nuclear factor-κB (NF-κB)-dependent signaling in the early stage of infection. Inhibition of the PI3-K/Akt or NF-κB pathway enhanced cell death but no effect was observed on virus replication or PRV gene expression. Transiently-expressed GFP- or His-tagged PRV Us3a and Us3b cDNA protect cells against PRV-, avian reovirus- or bovine ephemeral fever virus-induced apoptosis in the cell lines. Us3a and Us3b transient over-expression upregulated several anti-apopototic signaling events, and the anti-apoptosis activity of Us3a is greater than that of Us3b. Kinase activity-deficient point or double point mutated Us3a lost the kinase activity of Us3a, which showed that kinase activity is required for the anti-apoptosis effect of Us3. Akt and NF-κB activation still occurred in UV-inactivated PRV- and cycloheximide-treated cells. In vivo study showed that PRV-infected trigeminal ganglion increases the expression of anti-apoptosis signaling molecules, including Akt, PDK-1 and IκBα, which is a similar result to that seen in the in vitro experiments. Our study suggests that signaling mechanisms may play important roles in PRV pathogenesis.
authors.affiliation: Department of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan.
authors.forename: Ching-Dong
authors.initials: CD
authors.lastname: Chang
authors.forename: Ping-Yuan
authors.initials: PY
authors.lastname: Lin
authors.forename: Ming-Huei
authors.initials: MH
authors.lastname: Liao
authors.forename: Chi-I
authors.initials: CI
authors.lastname: Chang
authors.forename: Jue-Liang
authors.initials: JL
authors.lastname: Hsu
authors.forename: Feng-Ling
authors.initials: FL
authors.lastname: Yu
authors.forename: Hung-Yi
authors.initials: HY
authors.lastname: Wu
authors.forename: Wen-Ling
authors.initials: WL
authors.lastname: Shih
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002460:Cell Line; D002867:Chromones / Q000494:pharmacology; D004249:DNA Damage; D004121:Dimethyl Sulfoxide; D005419:Flavonoids / Q000494:pharmacology; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000378:metabolism*; D009025:Morpholines / Q000494:pharmacology; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism*; D011557:Pseudorabies / Q000821:virology; D015398:Signal Transduction; D013552:Swine; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D002867:Chromones; D004121:Dimethyl Sulfoxide; D005419:Flavonoids; D009025:Morpholines; D016328:NF-kappa B; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt; D014764:Viral Proteins; D000077191:Wortmannin; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
keywords: Akt; Apoptosis; NF-κB; PRV; Us3