Membrane permeabilization by small hydrophobic nonstructural proteins of Japanese encephalitis virus

Yu Shiu Chang, Ching Len Liao, Chang Huei Tsao, Mei Chieh Chen, Chiu I. Liu, Li Kuang Chen, Yi Ling Lin

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

Infection with Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, may cause acute encephalitis in humans and induce severe cytopathic effects in various types of cultured cells. We observed that JEV replication rendered infected baby hamster kidney (BHK-21) cells sensitive to the translational inhibitor hygromycin B or α-sarcine, to which mock- infected cells were insensitive. However, little is known about whether any JEV nonstructural (NS) proteins contribute to virus-induced changes in membrane permeability. Using an inducible Escherichia coli system, we investigated which parts of JEV NS1 to NS4 are capable of modifying membrane penetrability. We found that overexpression of NS2B-NS3, the JEV protease, permeabilized bacterial cells to hygromycin B whereas NS1 expression failed to do so. When expressed separately, NS2B alone, but not NS3, was sufficient to alter bacterial membrane permeability. Similarly, expression of NS4A or NS4B also rendered bacteria susceptible to hygromycin B inhibition. Examination of the effect of NS1 to NS4 expression on bacterial growth rate showed that NS2B exhibited the greatest inhibitory capability, followed by a modest repression from NS2A and NS4A, whereas NS1, NS3, and NS4B had only trivial influence with respect to the vector control. Furthermore, when cotransfected with a reporter gene luciferase or β-galactosidase, transient expression of NS2A, NS2B, and NS4B markedly reduced the reporter activity in BHK-21 cells. Together, our results suggest that upon JEV infection, these four small hydrophobic NS proteins have various modification effects on host cell membrane permeability, thereby contributing in part to virus-induced cytopathic effects in infected cells.

Original languageEnglish
Pages (from-to)6257-6264
Number of pages8
JournalJournal of Virology
Volume73
Issue number8
Publication statusPublished - 1999
Externally publishedYes

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Japanese Encephalitis Virus
Japanese encephalitis virus
Hygromycin B
hygromycin B
Membranes
membrane permeability
Proteins
cytopathogenicity
proteins
cells
Permeability
Galactosidases
Cell Membrane Permeability
Viruses
galactosidases
Flavivirus
viruses
vector control
Encephalitis
luciferase

ASJC Scopus subject areas

  • Immunology

Cite this

Chang, Y. S., Liao, C. L., Tsao, C. H., Chen, M. C., Liu, C. I., Chen, L. K., & Lin, Y. L. (1999). Membrane permeabilization by small hydrophobic nonstructural proteins of Japanese encephalitis virus. Journal of Virology, 73(8), 6257-6264.

Membrane permeabilization by small hydrophobic nonstructural proteins of Japanese encephalitis virus. / Chang, Yu Shiu; Liao, Ching Len; Tsao, Chang Huei; Chen, Mei Chieh; Liu, Chiu I.; Chen, Li Kuang; Lin, Yi Ling.

In: Journal of Virology, Vol. 73, No. 8, 1999, p. 6257-6264.

Research output: Contribution to journalArticle

Chang, YS, Liao, CL, Tsao, CH, Chen, MC, Liu, CI, Chen, LK & Lin, YL 1999, 'Membrane permeabilization by small hydrophobic nonstructural proteins of Japanese encephalitis virus', Journal of Virology, vol. 73, no. 8, pp. 6257-6264.
Chang, Yu Shiu ; Liao, Ching Len ; Tsao, Chang Huei ; Chen, Mei Chieh ; Liu, Chiu I. ; Chen, Li Kuang ; Lin, Yi Ling. / Membrane permeabilization by small hydrophobic nonstructural proteins of Japanese encephalitis virus. In: Journal of Virology. 1999 ; Vol. 73, No. 8. pp. 6257-6264.
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