West Nile virus capsid protein inhibits autophagy by AMP-activated protein kinase degradation in neurological disease development

• Published in: PLoS pathogens Vol. 16; no. 1; p. e1008238
• Main Authors: Kobayashi, Shintaro, Yoshii, Kentaro, Phongphaew, Wallaya, Muto, Memi, Hirano, Minato, Orba, Yasuko, Sawa, Hirofumi, Kariwa, Hiroaki
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01.01.2020; Public Library of Science (PLoS)
• Subjects: Accumulation; Aggregates; Amino acids; AMP; AMP-activated protein kinase; AMP-Activated Protein Kinases - metabolism; Animals; Apoptosis; Autophagy; Biology and life sciences; Birds; Brain; C protein; Capsid protein; Capsid Proteins - physiology; Cell death; Cell Line, Tumor; Chlorocebus aethiops; Degradation; Disease; Encephalitis; Female; Funding; HEK293 Cells; HIV; Human immunodeficiency virus; Humans; Infections; Kinases; Laboratories; Medicine and health sciences; Mice, Inbred C57BL; Mutants; Mutation; Nerve Tissue Proteins - metabolism; Nervous System Diseases - enzymology; Nervous System Diseases - pathology; Nervous System Diseases - virology; Neuroblastoma; Neuroblastoma cells; Neuroblasts; Neurodegeneration; Neurological diseases; Neurons - metabolism; Neurons - virology; Pathogenesis; Phagocytosis; Plasmids; Protein Aggregation, Pathological; Proteins; Proteolysis; Public health; Replication; Research and Analysis Methods; Residues; Signs and symptoms; Software; Supervision; Ubiquitination; Vector-borne diseases; Vero Cells; Veterinary medicine; Viral infections; Viral Proteins - metabolism; Viruses; West Nile virus; West Nile virus - physiology; Zoonoses; Japan; Fiji
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1008238

West Nile virus (WNV) belongs to the Flaviviridae family and has emerged as a significant cause of viral encephalitis in birds and animals including humans. WNV replication directly induces neuronal injury, followed by neuronal cell death. We previously showed that accumulation of ubiquitinated protein aggregates was involved in neuronal cell death in the WNV-infected mouse brain. In this study, we attempted to elucidate the mechanisms of the accumulation of protein aggregates in the WNV-infected cells. To identify the viral factor inducing the accumulation of ubiquitinated proteins, intracellular accumulation of ubiquitinated proteins was examined in the cells expressing the viral protein. Expression of capsid (C) protein induced the accumulation, while mutations at residues L51 and A52 in C protein abrogated the accumulation. Wild-type (WT) or mutant WNV in which mutations were introduced into the residues was inoculated into human neuroblastoma cells. The expression levels of LC3-II, an autophagy-related protein, and AMP-activated protein kinase (AMPK), an autophagy inducer, were reduced in the cells infected with WT WNV, while the reduction was not observed in the cells infected with WNV with the mutations in C protein. Similarly, ubiquitination and degradation of AMPK were only observed in the cells infected with WT WNV. In the cells expressing C protein, AMPK was co-precipitated with C protein and mutations in L51 and A52 reduced the interaction. Although the viral replication was not affected, the accumulation of ubiquitinated proteins in brain and neurological symptoms were attenuated in the mouse inoculated with WNV with the mutations in C protein as compared with that with WT WNV. Taken together, ubiquitination and degradation of AMPK by C protein resulted in the inhibition of autophagy and the accumulation of protein aggregates, which contributes to the development of neurological disease.