The STRUCTURE AND FUNCTION OF THE REOVIRUS GENOME

Joklik, Wolfgang K.

doi:10.1111/j.1749-6632.1980.tb27961.x

Cited by 25 publications

(21 citation statements)

References 106 publications

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“…were present at the highest concentration. Proteins of the same size make up over 60 ~o of the orthoreovirus virion (Joklik, 1981). Another point of similarity was the presence of three size classes of dsRNA.…”

Section: Discussionmentioning

confidence: 99%

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Morphological and Biochemical Properties of Four Members of a Novel Group of Reoviruses Isolated from Aquatic Animals

Winton

Lannan

Fryer

et al. 1987

Journal of General Virology

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SUMMARYThe morphological, biochemical and growth characteristics of four members of the Reoviridae, three from the fish hosts, golden shiner (Notemigonus crysoleucas), chum salmon (Oncorhynchus keta) and channel catfish (Ictalurus punetatus) and one from American oyster (Crassostrea virginica), were compared. Electron microscopy of negatively stained virions revealed icosahedral particles approximately 75 nm in diameter composed of a double capsid. Complete particles had buoyant densities in CsC1 of 1.34 to 1.36 g/ml. The viruses replicated well in several fish cell lines, forming plaque-like syncytia in monolayer cultures. Each virus could be distinguished by the range of cell lines supporting its growth. Polyacrylamide gel electrophoresis showed that the genome of each virus was composed of 11 segments of dsRNA distributed among three size classes. There were three large, three medium and five small segments in each genome and each isolate had a unique electropherotype. The segments ranged from 2.5 × 10 6 to 0"31 X 106 mol. wt. with a total genome of approximately 15 × 10 6 mol. wt. Analysis by SDS-PAGE revealed that each virus had five major structural proteins. There were two large polypeptides of approximately 135 000 and 125 000 mol. wt., one medium size polypeptide of 70000 mol. wt. and two small polypeptides of 45000 and 34000 mol. wt. Of the major structural proteins, those of approximately 70000 and 34000 tool. wt. were consistently present in the highest concentrations.Minor virion proteins were detected but were not characterized. These four viruses, isolated from aquatic animals, were unlike viruses of the six established genera of the Reoviridae.

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Section: Discussionmentioning

confidence: 99%

“…were present in the highest concentrations. Polypeptides with these molecular weights are the major outer capsid proteins of mammalian reoviruses (Joklik, 1981). The two large proteins required extended electrophoresis (16 h, 10 mA) to achieve adequate separation, particularly for CRV.…”

Section: Eleetrophoresis Of Proteinsmentioning

confidence: 99%

Morphological and Biochemical Properties of Four Members of a Novel Group of Reoviruses Isolated from Aquatic Animals

Winton

Lannan

Fryer

et al. 1987

Journal of General Virology

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“…First, sensitivity to 55 °C in both laboratory strains (type 2 and type 3) is determined by the $4 gene (see above), although these two strains are genetically divergent (Joklik, 1980). Second, we have made a preliminary examination of some of these properties in a variety of reovirus field isolates.…”

Section: Inactivation By 33%o Ethanolmentioning

confidence: 99%

“…Sigma 3 is the virion component which is responsible for the inhibition of host cell RNA and protein synthesis (Sharpe & Fields, 1982). The M2 gene, encoding ~tlC, the outer capsid polypeptide that mediates the response to proteolysis (Joklik, 1972(Joklik, , 1980Mustoe et al, 1978b;Rubin & Fields, 1979) determines the response of reovirus to 1 ~ phenol or 33~ ethanol.…”

Section: Inactivation By 33%o Ethanolmentioning

confidence: 99%

Genetic Studies on the Mechanism of Chemical and Physical Inactivation of Reovirus

Drayna

Fields

1982

Journal of General Virology

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SUMMARYThe three serotypes of reovirus differ markedly in their response to a variety of chemical inactivating agents. We used intertypic recombinants containing various combinations of genes derived from the parental serotypes to study the basis of these differences. In addition to recombinants derived from types 1 and 3, and 2 and 3, we were able to isolate recombinants derived from types 1 and 2, suggesting that these two serotypes also undergo unrestricted reassortment. The intertypic recombinants behaved like one parent or the other in the presence of the inactivating agents and allowed us to determine the genes responsible for each difference. Recombinants derived from crosses between wild-type parental serotypes produced straightforward results, while recombinants derived from mutagenized, temperature-sensitive parents often did not. Sensitivity to 2.5 M-guanidine-HC1 and pH 11 was determined by the S1 gene, sensitivity to 55 °C and 1 ~o SDS was determined by the $4 gene, and sensitivity to 33~ ethanol and to 1 ~ phenol was determined by the M2 gene. Thus, relatively nonspecific chemical agents appear to have their predominant effect on specific proteins of the reovirus virion. INTRODUCTIONThe mammalian reoviruses are classified as enteric viruses by their site of isolation and by their chemical and physical properties. These properties include resistance to proteolytic enzymes, acid, detergents and heat (Bitton, 1980). The biochemical and structural properties that allow the reoviruses, and other enteric viruses, to resist inactivation by these agents are not known. The genetic organization of mammalian reoviruses presents a unique opportunity to gain insight into the nature of the virus components responsible for determining the responses to such treatments. The three serotypes of human reoviruses are able to reassort dsRNA gene segments in mixed infection to produce recombinants (reassortants). These recombinants carry various combinations of discrete genes derived from the two parental serotypes. When two serotypes of reovirus differ in a biological or structural property, we have shown that intertypic recombinants can segregate the difference and thereby allow us to identify the gene(s) which specifies the difference (Fields, 1981). In this report, we show that the three reovirus serotypes differ in their sensitivity to detergents, heat, high pH and several organic denaturants. Intertypic recombinants segregate these differences and have allowed us to determine the genes responsible.

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“…After electrophoresis for RNA, the gel was fixed in 50% methanol and 12% acetic acid for 3 min. The SDS in the gel was removed (JOKLIK, 1981(JOKLIK, , 1983a1983b;WINTON, et al , 1987). The RNA pattern of LSV was closer to that of CSV, but quite different.…”

Section: Silver Stainmentioning

confidence: 99%

Characteristics of a new reo-like virus isolated from landlocked salmon (Oncorhynchus masou Brevoort).

Hsu¹,

Chen²,

Wu³

1989

Fish Pathol.

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The STRUCTURE AND FUNCTION OF THE REOVIRUS GENOME

Cited by 25 publications

References 106 publications

Morphological and Biochemical Properties of Four Members of a Novel Group of Reoviruses Isolated from Aquatic Animals

Morphological and Biochemical Properties of Four Members of a Novel Group of Reoviruses Isolated from Aquatic Animals

Genetic Studies on the Mechanism of Chemical and Physical Inactivation of Reovirus

Characteristics of a new reo-like virus isolated from landlocked salmon (Oncorhynchus masou Brevoort).

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