Synthesis of Seoul virus RNA and structural proteins in cultured cells.
Seoul virus is a hantavirus that causes hemorrhagic fever with renal syndrome (HFRS). The virion has a tripartite (S, M, and L) negative-stranded RNA genome, which is characteristic of the family Bunyaviridae. However, the molecular basis of virus replication is not well known. We established a Northern blot hybridization (NB) procedure using digoxygenin-labeled RNA probes, to quantitate the hantaviral plus- and minus-strand RNAs separately. Virus RNA replication was analyzed in infected Vero E6 cells. When the Vero E6 cells were infected with Seoul virus strain KI-83-262 (KI) at m.o.i. = 0.25, the plus-strand RNA was detected within 1 h post-infection (hpi), and the minus-strand RNA was detected subsequently. Using laser confocal microscopy, the nucleocapsid protein (NP) was detected within 2 hpi, and accumulated as scattered granules in the cytoplasm until 24 hpi. In contrast, the G2 protein first appeared at 8 hpi, was immediately transported to the Golgi, and accumulated in the Golgi until 24 hpi. Infectious virus particles were released into the medium at 24 h hpi. These findings indicate that hantavirus RNA replication starts with the appearance of NP at 2 hpi, glycoproteins then accumulate gradually in the Golgi, and virion formation is initiated once the viral RNAs and proteins have accumulated.
D000818:Animals; D002522:Chlorocebus aethiops; D006023:Glycoproteins / Q000096:biosynthesis; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000096:biosynthesis; D015347:RNA Probes; D012367:RNA, Viral / Q000096:biosynthesis*; D029263:Seoul virus / Q000502:physiology*; D014709:Vero Cells; D014758:Viral Core Proteins / Q000096:biosynthesis; D014764:Viral Proteins / Q000096:biosynthesis*; D014779:Virus Replication
D006023:Glycoproteins; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D015347:RNA Probes; D012367:RNA, Viral; D014758:Viral Core Proteins; D014764:Viral Proteins