Hantaan Virus Replication Is Promoted Via AKT Activated Mitochondria OXPHOS

【Background】China is one of the most severely affected epidemic countries of Hemorrhagic Fever with Renal Syndrome（HFRS）in the world,Hantaan virus（HTNV）is the major pathogen of HFRS in China.However,there are no specific therapeutic drugs against HTNV infection by now.Virus replication relies on energy and resource substance from host cells.Glycolysis and oxidative phosphorylation（OXPHOS）are two major metabolic pathways that produce energy in eukaryotic cells,which can be exploited by virus during its replication in host cells.Mechanism of how HTNV utilizing metabolic pathways in host cells after infection remains unknown.【Objectives】This research meant to elucidate the major metabolic pathway exploited by HTNV during its replication in host cells and its underlying molecular mechanism,this research may enrich our acknowledge about HTNV biological characteristics and pathogenesis.【Methods and Results】1.HTNV preferred to exploit mitochondrial OXPHOS during its replication in host cells,rather than glycolysis pathway;Mitochondrial OXPHOS or aerobic glycolysis changes after HTNV infection were tested by Seahorse XF Analyzers in different cells,which showed that HTNV replicaton can increase mitochondrial OXPHOS levels in different cells,however,there is no consistent trends of glycolysis capacity changes in different cells after HTNV infection.Meanwhile,OXPHOS level increase caused by HTNV replication did not vanish when infection period prolonging.2.HTNV can promote mitochondrial biosynthesis;We found that mitochondrial biosynthesis increased explicitly after HTNV infection via western blot,RT-q PCR and BN-PAGE.3.HTNV replication induced AKT phosphorylation and p AKT-ser⁴⁷³translocation from cytoplasm to mitochondria,which leaded to mitochondrial OXPHOS level increase after HTNV infection;We found that p AKT-ser⁴⁷³partially translocated from cytoplasm to mitochondria after HTNV infection by detecting p AKT-ser⁴⁷³in mitochondria and cytoplasm respectively,Immunofluorescence Assay results also verified that.Such translocation induced by HTNV replication can be blocked by PI3K-AKT inhibitor BEZ235.BEZ235 treatment can also inhibit mitochondrial biosynthesis and OXPHOS level increase caused by HTNV replication.4.Exploring the mechanism of mitochondrial OXPHOS increase caused by AKT activation.RXRXXS/T is an AKT substrate motif,by screening amino acid sequences of mitochondria proteins,we found that PNPT contained two RXRXXS/T sequences which can be phosphorylated by AKT.Furthermore,we verified that RXRXXS/T motif in PNPT can be phosphorylated by AKT through Co-IP assay,the phosphorylation level of PNPT RXRXXS/T motif increased after HTNV infection.Moreover,PNPT can interact with p AKT-ser⁴⁷³after HTNV infection.【Conclusion】HTNV exploited mitochondrial OXPHOS to provide energy for its replication in host cells.By activating AKT and inducing p AKT-ser⁴⁷³translocation from cytoplasm to mitochondria,HTNV replication can promote mitochondrial biosynthesis and mitochondrial OXPHOS.PNPT,which can mediate mitochondrial RNA importing and decaying,may be a substrate of mitochondrial p AKT-ser⁴⁷³during HTNV replication in host cells.