Cytoplasmic Fractions Associated with Semliki Forest Virus Ribonucleic Acid Replication

Friedman, Robert M.; Berezesky, Irene K.

doi:10.1128/jvi.1.2.374-383.1967

Cited by 68 publications

(62 citation statements)

References 28 publications

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“…Numerous studies have shown that virus-specific RNA polymerase activity is associated with cytoplasmic membranes together with heterogenous RNA (25)(26)(27). The role of 26S RNA as messenger for the structural proteins was finally proven by its in vitro translation in the presence of ER membranes, which yields all structural proteins (28).…”

Section: Replication Of Alphavirus Rnasmentioning

confidence: 99%

Functions of alphavirus nonstructural proteins in RNA replication

Kääriäinen

Ahola

2002

Progress in Nucleic Acid Research and Molecular Biology

107

111

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Section: Replication Of Alphavirus Rnasmentioning

confidence: 99%

Functions of alphavirus nonstructural proteins in RNA replication

Kääriäinen

Ahola

2002

Progress in Nucleic Acid Research and Molecular Biology

107

111

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“…As in other positive-strand RNA viruses, the RCs of alphaviruses are associated with altered intracellular membranes, which were first described in the late 1960s and early 1970s (13,14,18). In these early studies, it was shown that virus replication induces bulb-shaped membrane invaginations with a diameter of ϳ50 nm, which were called spherules.…”

mentioning

confidence: 99%

Phosphatidylinositol 3-Kinase-, Actin-, and Microtubule-Dependent Transport of Semliki Forest Virus Replication Complexes from the Plasma Membrane to Modified Lysosomes

Spuul

Balistreri

Kääriäinen

et al. 2010

J Virol

163

217

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Like other positive-strand RNA viruses, alphaviruses replicate their genomes in association with modified intracellular membranes. Alphavirus replication sites consist of numerous bulb-shaped membrane invaginations (spherules), which contain the double-stranded replication intermediates. Time course studies with Semliki Forest virus (SFV)-infected cells were combined with live-cell imaging and electron microscopy to reveal that the replication complex spherules of SFV undergo an unprecedented large-scale movement between cellular compartments. The spherules first accumulated at the plasma membrane and were then internalized using an endocytic process that required a functional actin-myosin network, as shown by blebbistatin treatment. Wortmannin and other inhibitors indicated that the internalization of spherules also required the activity of phosphatidylinositol 3-kinase. The spherules therefore represent an unusual type of endocytic cargo. After endocytosis, spherule-containing vesicles were highly dynamic and had a neutral pH. These primary carriers fused with acidic endosomes and moved long distances on microtubules, in a manner prevented by nocodazole. The result of the large-scale migration was the formation of a very stable compartment, where the spherules were accumulated on the outer surfaces of unusually large and static acidic vacuoles localized in the pericentriolar region. Our work highlights both fundamental similarities and important differences in the processes that lead to the modified membrane compartments in cells infected by distinct groups of positive-sense RNA viruses.All positive-strand RNA viruses replicate their genomes in association with cellular membranes. The formation and activity of the membrane-bound replication complexes (RCs) can result in extensive alteration of membrane structures (11,40,48). Different viruses use different cytoplasmic membrane compartments as platforms for replication. Currently, there is only a limited understanding of how the virus-encoded and cellular proteins coordinate the formation of the replicationinduced membrane structures. We address the mechanisms of membrane-bound replication with alphaviruses, particularly Semliki Forest virus (SFV). The alphaviruses comprise several human and animal pathogens, including the encephalitogenic alphaviruses (e.g., Western, Eastern, and Venezuelan equine encephalitis viruses) as well as the recently reemerging chikungunya virus, which belongs to the SFV clade of alphaviruses. During the past 5 years, chikungunya virus has caused more than 2 million infections and 500 deaths, and a new strain has spread throughout the areas surrounding the Indian Ocean (50). The alphaviruses use mosquitoes as intermediate hosts and transmission vectors, and at present no vaccines or antivirals are available to control these infections.The cytoplasmic replication of alphaviruses depends on the four viral nonstructural (ns) proteins, nsP1 to nsP4, which are all essential and act as a membrane-bound replication complex. The nsPs are tran...

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“…The synthesis of arbovirus-directed ribo nucleic acid has been studied recently in considerable detail with group A viruses. In the case of Semliki forest virus (SFV), three distinct types of RNA were demon strated during viral replication, which were 42S single-stranded, 26S ribonuclease-sensi tive and 20S double-stranded forms [2]. Similar observations have been presented with another group A member, Sindbis virus (SBV) [1].…”

mentioning

confidence: 58%

Replication and Synthesis of Japanese Encephalitis Virus Ribonucleic Acids in Vero Cells

Nishimura

Tsukeda

1971

Japanese Journal of Microbiology

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Neither replication nor ribonucleic acid synthesis of Japanese encephalitis virus (JEV) in Vero cells was affected by actinomycin D at a concentration of 0.5,g/ml, but both were markedly inhibited by formycin A, an inhibitor of RNA synthesis, at a concentra tion of between 10 and 25Synthesis of virus-specific RNA's was then studied in the presence of actinomycin D. The viral RNA synthesis, as estimated from the eventual virus yields in cells treated with formycin A at different intervals from infection, was found to proceed 6 to 7 hr in advance of mature virus formation which began at 10 hr after infection. The newly synthesized viral RNA detected by a 6-hr pulse-labeling with 3H-uridine was observed at the 43S region in a sucrose gradient centrifugation. With a shorter pulse, however, a rapidly labeled virus-specific RNA was detected first in the 10 to 15S region, which was soluble in 2 ivt NaCl, and this label moved to the ribo nuclease-resistant 26S RNA during a subsequent 6-hr period. Further studies of this 26S RNA by pulse-chase experiments demonstrated that a minor 20S double-stranded form accumulated as a by-product in the viral 43S RNA replication. Thus the two ribonuclease-resistant forms of virus-specific RNA, 26S and 20S, were identified in JEV infected Vero cells. Results of these experiments suggest that the 43S single-stranded viral RNA and the 20S double-stranded minor RNA might be formed through the intermediate type 26S RNA which is partially ribonuclease-resistant.

show abstract

Cytoplasmic Fractions Associated with Semliki Forest Virus Ribonucleic Acid Replication

Cited by 68 publications

References 28 publications

Functions of alphavirus nonstructural proteins in RNA replication

Functions of alphavirus nonstructural proteins in RNA replication

Phosphatidylinositol 3-Kinase-, Actin-, and Microtubule-Dependent Transport of Semliki Forest Virus Replication Complexes from the Plasma Membrane to Modified Lysosomes

Replication and Synthesis of Japanese Encephalitis Virus Ribonucleic Acids in Vero Cells

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