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Interferon-beta (IFN-beta) strongly inhibited the expression of the hemagglutinin-neuraminidase (HN) gene of Newcastle disease virus (NDV), a paramyxovirus, in HeLa cells under the conditions where it did not affect the expression of the four upstream genes encoding the nucleocapsid protein, phosphoprotein, membrane protein and fusion protein. Even the downstream gene, encoding the large protein as well as the genome replication, appeared to be less susceptible to IFN-beta than the HN gene. This selective action of IFN-beta did not appear to be attributable to its well characterized antiviral mechanisms such as acceleration of RNA decay and translation inhibition. No similar down-regulation of a particular gene expression was found with another paramyxovirus, Sendai virus, or with a rhabdovirus, vesicular stomatitis virus, or seems to have been reported previously with any negative-strand RNA viruses. This new effect of IFN-beta thus suggests gene expression mechanism unique to NDV and may further lead to the discovery of a novel biochemical effect of IFN-beta.

A rat fibroblast cell line, 3Y1 is nonpermissive for infection by several negative-strand-RNA viruses including influenza virus A, Sendai virus, Newcastle disease virus and vesicular stomatitis virus (VSV), but not refractory to that of a positive-strand-RNA virus, Sindbis virus. To elucidate the mechanism of the restricted viral growth, we compared the replication pattern of VSV in 3Y1 cells and in baby hamster kidney cells, which are fully permissive for those viruses. The results indicated that the restriction was imposed predominantly on the transcription of the viral RNA. The subsequent steps such as protein synthesis and nucleocapsid assembly appeared to occur normally, although these levels remained low due to the restricted transcription. Virus attachment onto, and penetration into, 3Y1 cells were also not restricted.

We examined the antiviral effects of three oligopeptides, carbobenzoxy (Z)-d-Phe-Ile-Gly, Z-d-Leu-Ile-Gly and Z-d-Phe-Phe-Gly, which mimic the N-terminal regions of F1 glycoproteins of two Newcastle disease virus strains (Miyadera and D26) and Sendai virus, respectively. Only one of these peptides, Z-d-Phe-Phe-Gly, significantly and with a similar potency inhibited viruses of homologous and heterologous F1 N-terminal sequences, suggesting no strict sequence requirement for inhibition. Furthermore, the enveloped RNA viruses of several different families showed essentially the same sensitivity to the three peptides as the paramyxoviruses, while a nonenveloped RNA virus was not susceptible to any of them. In addition, the Z-d-Phe-Phe-Gly peptides was effective only when the virus particles had been pretreated before infection.

Most of the information on the structure, function and replication of paramyxoviruses has been obtained from studies on Newcastle disease virus (NDV), simian virus 5 (SV5), and Sendal virus (see reviews by Kingsbury, 1973; Choppin and Compans, 1975; Rott and Klenk, 1976). The virion consists of a helical ribonucleoprotein, the nucleocapsid, which is surrounded by an envelope (Fig.l). The nucleocapsid is composed of the single stranded RNA genome and a carbohydrate-free polypetide subunit, the NP protein. The envelope contains lipid which is derived from the plasma membrane of the host cell (Klenk and Choppin, 1969) and is present in the form of a bilayer (Landsberger et al., 1971). Associated with the outer side of the bilayer are spikes which are composed of glycoproteins (Klenk, et al., 1970; Chen, et al., 1971), the inner side is coated by the carbohydrate-free M protein. There are 2 types of spikes: one type consists of glycoprotein HN which has hemagglutinating and neuraminidase activity (Scheid et al., 1972; Scheid and Choppin, 1973; Seto et al., 1973;Tozawa, et al., 1973; Shimizu, et al., 1974), and the other one consists of glycoprotein F which induces cell fusion and hemolysis (Homma and Ohuchi, 1973; Scheid and Choppin, 1974; Seto, et al., 1974). It is generally agreed that these activities reflect the essential roles which both

Purified virions of Newcastle disease virus (NDV) were found to contain protein kinase activity which was, like other virion-associated kinases, stimulated by Mg2+, and totally independent of Ca2+ and cAMP. The kinase phosphorylated preferentially the P and NP polypeptides of NDV. Triton-KCl fractionation of the virions has shown that the protein kinase activity may be associated with glycoprotein-free subviral particles, but not with nucleocapsids containing either only NP or some L and P proteins together with NP as protein constituent.

About 15 serial passages of wild type mumps virus (Sasazaki strain) in the amnion sac of chick embryo (CE) yielded a CE-adapted strain which was poorly replicative and did not form plaques in Vero cells where the wild strain grew well. In the course of this limited replication of the CE-adapted strain in Vero cells, we have analysed the viral protein and RNA synthesis. It was found that protein synthesis took place very efficiently at least early in infection by 12 h. The subsequent rate of synthesis remained, however, at a low level without showing the progressively increasing synthesis observed with the wild strain. Furthermore, 50S genomic RNA was synthesized early in the limited infection, but the subsequent synthesis was markedly suppressed. In addition, the other virus-specific RNA species could not be detected throughout. Thus the amplified RNA synthesis observed in the permissive CE cells and in the wild strain-infection of Vero cells seemed not to occur in the limited replication. Neither interferon nor DI (defective interfering) RNA was involved in the limited virus growth. When Vero cells were infected with the wild strain 6 to 8 h before inoculation of CE-adapted strain, growth restriction was overcome and the yield of the latter virus was greatly enhanced by a factor more than 10(3). These results suggest that through adaptation to CE, mumps virus may be altered in such a way that there is a restriction, probably at a step (s) involved in amplification of the viral RNA synthesis in Vero cells and that the restriction may be overcome by the simultaneous genome expression of the prototype wild strain.

Smooth membrane fractions were prepared from the cytoplasmic extract of BHK-21 cells infected with Newcastle disease virus (NDV). These membranes exhibited high hemagglutinating, neuraminidase, and hemolytic activity but little infectivity, suggesting that they might be precursors for viral envelope. When such membranes were adsorbed to the monolayers of uninfected BHK-21 cells at 4 degrees C and then incubated at elevated temperature for a couple of hours, the cells became highly hemadsorptive even in the presence of cycloheximide. This phenomenon occurred between 15 degrees C and 25 degrees C, and was maximal at 31 degrees C, where approximately 4 times more erythrocytes were adsorbed than to the cells incubated at 4 degrees C. Immunofluorescent staining suggested that diffusion of viral antigens might occurred rapidly over the entire surface of the cells. Cell fractions containing virions induced hemadsorption in uninfected cells, too. However, induction occurred now at 31 degrees C and was maximal at 37 degrees C, and erythrocytes appeared to be adsorbed not to the entire surface of the monolayer but restricted areas of the cells. The diffusion of viral antigens on the cell surface was not so significant under these conditions. On the basis of these findings the possible role of the membranes of the smooth endoplasmic reticulum in virus replication is discussed.