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Association of Virus Specific Replicative Ribonucleic Acid with Nuclear Membrane in Chick Embryo Cells Infected with Japanese Encephalitis Virus

Tomoyoshi Yasuda

Masa-Atsu Yamada

^* Department of Virology and Rickettsiology
Department of Technology, National Institute of Health in Tokyo, Kamiosaki 2-10-35, Shinagawa-ku, Tokyo 141
Faculty of Pharmaceutical Science. University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113, Japan

Using high resolution electron microscopic autoradiography and velocity sedimentation, RNA synthesis was examined in chick embryo cells infected with Japanese encephalitis virus (JEV). RNA was labelled with ³H-uridine for 5 min or 10 min at 15 h after infection in the presence of actinomycin D and D-glucosamine. Microautoradiography showed significant numbers of silver grains on the nuclear membranes of 5 min pulse-labelled thin cell sections.

The RNA species in membrane fractions obtained from the nucleus and cytoplasm of the infected cells were analysed by sucrose density gradient sedimentation. Radioactive 23S replicative form RNA and 8-12S RNA were obtained from the outer membrane fractions of the nuclear envelope. Labelled 42S virus RNA was obtained from the fractions containing large vesicle membranes and plasma membranes.

These results suggest that JEV-RNA synthesis is initiated in the perinuclear region in close association with the outer membranes of the nuclear envelope.

Received 26 April 1977; accepted 18 August 1977.

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