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Duck enteritis virus activates CaMKKβ-AMPK to trigger autophagy in duck embryo fibroblast cells via increased cytosolic calcium.

ABSTRACT: BACKGROUND:The results of our previous study showed that impaired cellular energy metabolism contributes to duck enteritis virus-induced autophagy via the 5`-adenosine monophosphate-activated protein kinase (AMPK)/tuberous sclerosis complex 2/mammalian target of rapamycin pathway in duck embryo fibroblast (DEF) cells. However, it remains unknown whether any other underlying mechanisms of AMPK activation are involved in autophagy induction. METHODS:The activity of CaMKKβ and AMPK in DEF cells infected with DEV were evaluated.The Effect of inhibitory activity of CaMKKβ on DEV-induced autophagy was investigated. In addtion to, the cytosolic calcium level in DEF cells infected with DEV were evaluated.The Effect of inhibitory cytosolic calcium level on DEV-induced autophagy was investigated. RESULTS:In this study, duck enteritis virus (DEV) infection activated CaMKKβ and its substrate molecule AMPK at 36, 48, and 60 h post-infection (hpi). STO-609, a CaMKKβ inhibitor, or CaMKKβ siRNA significantly inhibited the activation of DEV to AMPK, LC3I to LC3II transformation, and GFP-LC3 puncta distribution. In addition, inhibition of CaMKKβ activity also significantly reduced progeny DEV titer and gB protein expression. Besides, cytosolic calcium (Ca2+) was higher in DEV-infected cells than mock controls at 36, 48, and 60 hpi, respectively. Treatment of DEV-infected cells with 1,2-Bis (2-aminophenoxy) ethane-N, N, N', N-tetraacetic acid (BAPTA-AM) significantly reduced intracellular Ca2+ ion concentrations, as well as CaMKKβ and AMPK activities, and subsequent autophagy, in addition to viral protein synthesis and viral titer. CONCLUSIONS:These results showed that elevated [Ca2+]cyto-mediated activation of CaMKKβ managed the activation of AMPK, which then positively regulated autophagy, thereby providing further insight into DEV-host interactions.


PROVIDER: S-EPMC6090797 | BioStudies | 2018-01-01


REPOSITORIES: biostudies

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