Characterization of temperature-sensitive HVJ (Sendai virus) infection in Vero cells: inhibitory mechanism of viral production at 41 degrees.
In a previous study, it was found that the synthesis of hemagglutinating virus of Japan (HVJ; Sendai virus)-specific proteins was inhibited at the transcriptional level at 41 degrees in LLC-MK2 cells. During an investigation of the temperature sensitivity of HVJ production in other host cells, the synthesis of HVJ-specific proteins was recognized even at 41 degrees in Vero cells. Viral production, however, was not detected, indicating the inhibition of steps after the synthesis of viral proteins. Hemadsorption activity was not detected at 41 degrees, suggesting problems with the envelope proteins, especially hemagglutinin-neuraminidase (HN) protein, at the cell membrane. Immunofluorescent staining and surface immunoprecipitation showed that HN protein was not present on the surface in spite of its localization in the cytoplasm. Further, analysis of the cell membrane fraction suggested that fusion (F) protein was integrated into the cell membrane but HN protein was not at 41 degrees. Electron microscopic observation showed that budding sites with spike structures formed and nucleocapsids assembled under the sites at 41 degrees without HN protein, although budded HVJ virions were not detected. At this time, F protein was exposed to the cell membrane and interacted with matrix and nucleocapsid proteins. The results suggested that the suppression of HVJ production at 41 degrees was due to the absence of HN protein in the membrane of Vero cells.
Hirayama
Etsuko
E
Institute of Molecular and Cellular Biology for Pharmaceutical Sciences, Kyoto Pharmaceutical University, Kyoto, Japan.
Ishida
Yo-ichi
Y
Sugimoto
Masao
M
Hiraki
Akihiro
A
Kim
Jeman
J
D000818:Animals; D002462:Cell Membrane / Q000821:virology; D002522:Chlorocebus aethiops; D015707:HN Protein / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008854:Microscopy, Electron; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000378:metabolism; D010253:Respirovirus Infections / Q000209:etiology / Q000473:pathology / Q000821:virology; D029082:Sendai virus / Q000472:pathogenicity* / Q000502:physiology / Q000648:ultrastructure; D013696:Temperature; D014709:Vero Cells; D014758:Viral Core Proteins / Q000378:metabolism; D014760:Viral Fusion Proteins / Q000378:metabolism; D014763:Viral Matrix Proteins / Q000378:metabolism; D019065:Virus Assembly; D014779:Virus Replication
D015707:HN Protein; C097020:M protein, Sendai virus; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D014758:Viral Core Proteins; D014760:Viral Fusion Proteins; D014763:Viral Matrix Proteins