Nuclear localization of Japanese encephalitis virus core protein enhances viral replication.
Japanese encephalitis virus (JEV) core protein was detected in both the nucleoli and cytoplasm of mammalian and insect cell lines infected with JEV or transfected with the expression plasmid of the core protein. Mutation analysis revealed that Gly(42) and Pro(43) in the core protein are essential for the nuclear and nucleolar localization. A mutant M4243 virus in which both Gly(42) and Pro(43) were replaced by Ala was recovered by plasmid-based reverse genetics. In C6/36 mosquito cells, the M4243 virus exhibited RNA replication and protein synthesis comparable to wild-type JEV, whereas propagation in Vero cells was impaired. The mutant core protein was detected in the cytoplasm but not in the nucleus of either C6/36 or Vero cell lines infected with the M4243 virus. The impaired propagation of M4243 in mammalian cells was recovered by the expression of wild-type core protein in trans but not by that of the mutant core protein. Although M4243 mutant virus exhibited a high level of neurovirulence comparable to wild-type JEV in spite of the approximately 100-fold-lower viral propagation after intracerebral inoculation to 3-week-old mice of strain Jcl:ICR, no virus was recovered from the brain after intraperitoneal inoculation of the mutant. These results indicate that nuclear localization of JEV core protein plays crucial roles not only in the replication in mammalian cells in vitro but also in the pathogenesis of encephalitis induced by JEV in vivo.
Mori
Yoshio
Y
Research Center for Emerging Infectious Diseases, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan.
Okabayashi
Tamaki
T
Yamashita
Tetsuo
T
Zhao
Zijiang
Z
Wakita
Takaji
T
Yasui
Kotaro
K
Hasebe
Futoshi
F
Tadano
Masayuki
M
Konishi
Eiji
E
Moriishi
Kohji
K
Matsuura
Yoshiharu
Y
D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001921:Brain / Q000821:virology; D002466:Cell Nucleolus / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D009033:Culicidae; D003593:Cytoplasm / Q000378:metabolism; D004664:Encephalitis Virus, Japanese / Q000254:growth & development / Q000502:physiology*; D004672:Encephalitis, Japanese / Q000821:virology; D005260:Female; D005816:Genetic Complementation Test; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D012313:RNA / Q000096:biosynthesis; D014709:Vero Cells; D014758:Viral Core Proteins / Q000737:chemistry / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis; D014779:Virus Replication
D014758:Viral Core Proteins; D014764:Viral Proteins; D012313:RNA
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