Structural components of Uukuniemi virus, a noncubical tick-borne arbovirus

Pettersson, Ralf F.; Kääriäinen, Leevi; Bonsdorff, Carl-Henrik von; Oker-Blom, N

doi:10.1016/0042-6822(71)90074-2

Cited by 50 publications

(54 citation statements)

References 16 publications

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“…This virus contains two envelope glycoproteins, Gl (Mr 70000) and G2 (Mr 65000) (Pettersson et al, 1971 ;yon Bonsdorff & Pettersson, 1975), the glycan structures of which have recently been determined (Pesonen et al, 1982). The unglycosylated Gl and G2 are about the same size (Mr 54000) (E. Kuismanen, unpublished results).…”

Section: Introductionmentioning

confidence: 99%

“…(p110) precursor, which is cotranslationally cleaved in the middle (Ulmanen et al, 1981). The nucleocapsid consists of three circular single-stranded RNA species L (Mr 2"4 × 106), M (Mr l'l x 106) and S (Mr 0.5 x l06) (Hewlett et al, 1977;Pettersson et al, 1977), multiple copies of the N protein (Mr 25000) (Pettersson et al, 1971) and a few copies of the L protein (Mr about 200000) (R. F. Pettersson, unpublished results).…”

Section: Introductionmentioning

confidence: 99%

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Characterization of Two Recombination-Complementation Groups of Uukuniemi Virus Temperature-sensitive Mutants

Gahmberg

1984

Journal of General Virology

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SUMMARYWith the aim of isolating temperature-sensitive (ts) mutants defective in virus maturation or glycoprotein transport, Uukuniemi virus, a bunyavirus, was mutagenized with N-methyl-N'-nitro-N-nitrosoguanidine. Out of 13 initial clones unable to grow at 39 °C (non-permissive temperature), five mutants which grew to titres above 107 p.f.u./ml at 32 °C (permissive temperature) were selected for further studies. The mutants fell into two coinciding recombination-complementation groups. Three group I mutants (ts7, 8 and 12) and two group II mutants (ts6 and 11) synthesized all three RNA segments and were able to form the corresponding nucleoproteins at 39 °C. Thus, members of these two recombination groups had a RNA-positive phenotype. All five mutants showed immunofluorescence when cells were stained at 39 °C using a doublestaining technique employing monoclonal antibodies against the glycoproteins G 1 or G2, and polyclonal antibodies against the nucleoprotein, N. We have previously shown that in cells infected with wild-type virus both the G1/G2 and the N proteins accumulate in the Golgi complex, the site of virus maturation. In cells infected with tsl2, accumulation of G1 and G2, but not N protein, was observed in the Golgi complex at 39 °C. The N protein was found evenly scattered in the cytoplasm, suggesting lack of interaction between the G1/G2 and N proteins. With ts6 and 11, G1 and G2 appeared to accumulate and aggregate in the endoplasmic reticulum (ER) at 39 °C. The location of the N protein coincided with that of the aggregated glycoproteins, suggesting that the N protein interacted with G 1/G2 already in the ER.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Characterization of Two Recombination-Complementation Groups of Uukuniemi Virus Temperature-sensitive Mutants

Gahmberg

1984

Journal of General Virology

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“…The antigens were localized by indirect immunofluorescence and immunoelectron microscopy with highly specific antisera directed against the structural proteins of one particular bunyavirus, Uukuniemi virus, the prototype of the Uukuvirus genus of Bunyaviridae (3). This virus consists of a lipoprotein membrane containing two envelope glycoproteins, Gl and G2 (Mr, about 70,000 and 65,000, respectively [27,36]), and a nucleocapsid, containing three single-stranded RNA segments (L, M, and S [25,28]), to which multiple copies of the N protein (Mr, 25,000) and a few copies of the L protein (Mr, 200,000) are associated (27). The two glycoproteins, which are encoded by the M RNA segment, are synthesized as a precursor (p110), which is cleaved cotranslationally roughly in the middle to yield Gl (34).…”

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confidence: 99%

“…Uukuniemi virus (4 mg), purified as described previously (26), was mixed with [35S]methionine-labeled virus. The virus was disrupted with 2% (vol/vol) Triton X-100 at 20°C for 10 min, and the nucleocapsid and envelope fractions were separated on CsCl gradients as described previously (25,27). Centrifugation was for 20 h at 45,000 rpm and 4°C in a Spinco SW50.1 rotor.…”

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confidence: 99%

Uukuniemi virus maturation: accumulation of virus particles and viral antigens in the Golgi complex.

Kuismanen¹,

Hedman²,

Saraste³

et al. 1982

Mol. Cell. Biol.

102

106

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We studied the maturation of Uukuniemi virus and the localization of the viral surface glycoproteins and nucleocapsid protein in infected cells by electron microscopy, indirect immunofluorescence, and immunoelectron microscopy with specific antisera prepared in rabbits against the two glycoproteins Gl and G2 and the nucleocapsid protein N. Electron microscopy of thin sections from infected cells showed virus particles maturing at smooth-surfaced membranes close to the nucleus. Localization of the G1/G2 and N proteins by indirect immunofluorescence at different stages after infection showed the antigens to be present throughout the cell interior but concentrated in the juxtanuclear region. The Gi/ G2 antiserum also appeared to stain the nuclear and plasma membranes. Double staining with tetramethylrhodamine isothiocyanate-conjugated wheat germ agglutinin, which preferentially stains the Golgi complex, and fluorescein isothiocyanate-conjugated anti-rabbit immunoglobulin G, which stained the G1/G2 or N proteins, showed that the staining of the juxtanuclear region coincided. Similarly, double staining for thiamine pyrophosphatase, an enzyme activity specific for the Golgi complex, showed the fluorescence and the cytochemical stain to coincide in the juxtanuclear region. Immunoperoxidase electron microscopy of cells permeabilized with saponin revealed that the viral glycoproteins were present in the rough endoplasmic reticulum and the nuclear and Golgi membranes; the latter was heavily stained. With this method, the N protein was localized to the cytoplasm, especially around smooth-surfaced vesicles in the Golgi region. Taken together, the results indicate that Uukuniemi virus and its structural proteins accumulate in the Golgi complex, supporting the idea that this compartment rather than the plasma membrane is the site of virus maturation. This raises the interesting possibility that deficient transport of the glycoproteins to the plasma membrane and hence their accumulation in the Golgi complex determines the site of virus maturation.A common feature of the viruses belonging to the large Bunyaviridae family (3) is that they appear to mature intracellularly by a budding process at the smooth-surfaced vesicles in the Golgi region (2,18,21,37). If this finding is indeed true, the maturation of the bunyaviruses differs from that of other enveloped viruses, which receive their envelopes either by budding at the plasma membrane (e.g., alpha-, myxo-, paramyxo-, and rhabdoviruses [4,12,13]), nuclear membrane (e.g., herpesviruses [5]), or the rough endoplasmic reticulum (e.g., coronaviruses [20]). The factors determining the site of maturation of enveloped viruses have remained largely unknown.

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“…The Uukuvirus genus comprises a single serogroup containing seven viruses, and in nature uukuviruses have been isolated from ticks, birds and certain rodents (Bishop & Shope, 1979;Bishop et al, 1980). The prototype of the genus is Uukuniemi virus (Oker-Blom et al, 1964) which has been the subject of many biochemical analyses (Gahmberg, 1984;Kuismanen, 1984;Kuismanen et al, 1982Kuismanen et al, , 1984Parker & Hewlett, 1981 ;Pesonen et aL, 1982;Pettersson & K/ifiri~inen, 1973 ;Pettersson et al, 1971Pettersson et al, , 1977von Bonsdorff & Pettersson, 1975;Ulmanen et al, 1981). The other six recognized members of the genus, Grand Arbaud, Manawa, Oceanside, Ponteves, Zaliv-Terpeniya and EgAn1825-61, are serologically related to Uukuniemi virus but no structural or biochemical analyses of these viruses have been reported.…”

Section: Introductionmentioning

confidence: 99%