The proteins of Japanese encephalitis virus

SUMMARYA mouse monoclonal antibody (MAb) to dengue 4 (DEN-4) virus reacted with the antigen in the nucleus as well as in the cytoplasm of DEN-4-infected mammalian and mosquito cells, as demonstrated by the peroxidase-antiperoxidase staining method. The intranuclear antigen appeared to accumulate at the nucleoli, forming spots, whereas the cytoplasmic antigen appeared to be localized mainly in large perinuclear loci in the infected cells. The MAb-reactive antigen was produced in the presence of actinomycin D, which caused the accumulation in the nucleus to be altered to a dispersed pattern. Radioimmunoprecipitation analysis of [35S]methionine-labelled purified virions and Western blot analysis of the antigens prepared from the infected mammalian and mosquito cells showed that the MAb was directed against the DEN-4 virus core protein (M~ 15-5K). These results indicated that the DEN-4 virus core protein was partially transported, soon after its synthesis in the cytoplasm, into the nucleus and accumulated at the nucleoli.

Section: Effect Of Actinomycin D On the Accumulation Of Intranuclear mentioning

confidence: 99%

Detection of Dengue 4 Virus Core Protein in the Nucleus. I. A Monoclonal Antibody to Dengue 4 Virus Reacts with the Antigen in the Nucleus and Cytoplasm

Tadano

Makino

Fukunaga

et al. 1989

“…The glycoprotein GP 19 has been labelled with radioactive carbohydrate or amino acids and detected in slowly sedimenting haemagglutinating particles and in some preparations of virions (Stollar, 1969;Shapiro et al, 1971;Trent & Qureshi, 1971;Shapiro et al, 1972;Westaway, 1973Westaway, , 1975Boulton & Westaway, 1976). Since the analyses presented in this (Wright et al, 1980).…”

Section: Gp19 Associated With Purified Virusmentioning

confidence: 99%

“…Envelope protein (E) and a membrane-like protein (M) are found in both virions and slowly sedimenting haemagglutinating particles, core protein (C) is found only in virions, and a fourth polypeptide, a glycoprotein (GP 19), is detected both in slowly sedimenting haemagglutinating particles and under some conditions in virions (Stollar, 1969;Shapiro et al, 1971;Trent & Qureshi, 1971;Shapiro et al, 1972;Westaway, 1973Westaway, , 1975Boulton & Westaway, 1976). We are currently investigating the synthesis of Kunjin virus-specified glycoproteins in infected cells, and comparing the properties of E and GPI9 from virions and slowly sedimenting haemagglutinating particles with those of their likely precursors in infected cells (Wright et al, 1980(Wright et al, , 1981.…”

Section: Introductionmentioning

confidence: 99%

Envelope Protein of the Flavivirus Kunjin is Apparently Not Glycosylated

Wright

1982

SUMMARYThe envelope protein E (formerly designated V3) of the flavivirus Kunjin was not labelled with radioactive galactose, mannose or glucosamine during virus growth in Vero cells. On electrophoresis through polyacrylamide gels containing SDS, the envelope (E) protein migrated more rapidly than related intracellular virus-specified glycoproteins. Furthermore, E had a density in CsC1 solution consistent with that of a protein lacking carbohydrate, and did not bind to concanavalin A-agarose. In contrast, the envelope glycoprotein of Murray Valley encephalitis virus (MVEV) did bind to concanavalin A under similar conditions and was readily labelled with radioactive mannose. These results suggested that the E protein of Kunjin virus was not glycosylated, a feature not shared with MVEV and West Nile virus (WNV), whose properties were consistent with the presence of oligosaccharides attached to the envelope proteins. When Kunjin virions were labelled with radioactive glucosamine, the label was contained in GP19 (formerly NV2). The glycopeptides derived by Pronase digestion of GP19 from Kunjin virions were larger than those derived from GP 19 obtained from infected cells.

“…PrM is the precursor of the membrane protein M. N-terminal amino acid analyses of envelope protein (E), core protein (C) and M, and of the non-structural (ns) proteins NS1, NS3 and NS5 established their cleavage sites in the YF virus polyprotein (Rice et al, 1985;Bell et al, 1985). NS1, NS3 and NS5 correspond to the ns proteins originally described as NV3, NV4 and NV5 respectively (Shapiro et al, 1971 ;Westaway, 1973).…”

Section: Introductionmentioning

confidence: 99%

Gene Mapping and Positive Identification of the Non-structural Proteins NS2A, NS2B, NS3, NS4B and NS5 of the Flavivirus Kunjin and Their Cleavage Sites

Speight

Coia

Parker

et al. 1988

115

SUMMARYPartial N-terminal amino acid analyses of five radiolabelled non-structural (ns) proteins specified by Kunjin (KUN) virus provided positive identification of NS3, NS5 and three previously hypothetical ns proteins of flaviviruses, ns2a, ns2b and ns4b. Their correct gene order was obtained from their deduced amino acid sequences. Thus the gene order for KUN virus relative to that proposed for yellow fever (YF) virus was as follows: KUN 5'...GP44-P19.P10.P71.(?)-P21-P98-3', YF 5'...NSl.ns2a.ns2b.NS3.ns4a-ns4b.NS5-3'. The identity of GP44 as NS1 was assumed from the known nucleotide and deduced amino acid sequences; ns4a was not identified. The cleavage sites in the polyprotein for KUN NS2B, NS3 and NS5 were identical, Lys-Arg~Gly, similar in form to the sequence Arg-Arg~Ser defined at the cleavage sites ofYF NS3 and NSS. A new consensus cleavage site for NS1, NS2A and NS4B in the form VaI-X-Ala~, where X is any one of several uncharged amino acids, was found at corresponding sites homologous to those of KUN virus in all published flav ivirus sequences (a total of 18 sites). N S 1 and N S4B, but not N S2A, were preceded by a putative signal sequence.