The Spread of
            <i>Rice Dwarf Virus</i>
            among Cells of Its Insect Vector Exploits Virus-Induced Tubular Structures

Wèi, Tàiyún; Kikuchi, Akira; Moriyasu, Yusuke; Suzuki, Nobuhiro; Shimizu, Takumi; Hagiwara, Kyoji; Chen, Hongyan; Takahashi, M.; Ichiki-Uehara, Tamaki; Omura, Toshihiro

doi:10.1128/jvi.00537-06

Cited by 93 publications

(132 citation statements)

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“…The antibodies against viral particles were conjugated directly to FITC as described (39). Rabbit polyclonal anti-VSVG was a kind gift from I. M. Jones (Reading University, Reading, U.K.).…”

Section: Methodsmentioning

confidence: 99%

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The P2 capsid protein of the nonenveloped rice dwarf phytoreovirus induces membrane fusion in insect host cells

Zhou

Wei

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Insect transmission is an essential process of infection for numerous plant and animal viruses. How an insect-transmissible plant virus enters an insect cell to initiate the infection cycle is poorly understood, especially for nonenveloped plant and animal viruses. The capsid protein P2 of rice dwarf virus (RDV), which is nonenveloped, is necessary for insect transmission. Here, we present evidence that P2 shares structural features with membrane-fusogenic proteins encoded by enveloped animal viruses. When RDV P2 was ectopically expressed and displayed on the surface of insect Spodoptera frugiperda cells, it induced membrane fusion characterized by syncytium formation at low pH. Mutational analyses identified the N-terminal and a heptad repeat as being critical for the membrane fusion-inducing activity. These results are corroborated with results from RDV-infected cells of the insect vector leafhopper. We propose that the RDV P2-induced membrane fusion plays a critical role in viral entry into insect cells. Our report that a plant viral protein can induce membrane fusion has broad significance in studying the mechanisms of virus entry into insect cells and insect transmission of nonenveloped plant and animal viruses.

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Section: Methodsmentioning

confidence: 99%

“…After a 2-h adsorption period at 25°C, the inoculum was removed. After 5 days incubation at 25°C, VCMs were fixed by 2% paraformaldehyde, stained with viral particle-specific IgG conjugated to FITC (39), and visualized by fluorescence microscopy. Mockinoculated healthy cells were processed in the same manner.…”

Section: Pcr Primer Sequencesmentioning

confidence: 99%

The P2 capsid protein of the nonenveloped rice dwarf phytoreovirus induces membrane fusion in insect host cells

Zhou

Wei

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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“…This formation is intrinsic to viral particle functioning in virus-infected plants and insect cells [1][2][3]. Southern rice black-streaked dwarf virus (SRBSDV), a newly discovered member of the genus fijivirus [4,5], causes the devastating southern rice black-streaked dwarf and maize rough dwarf diseases in crops, which lead to severe yield losses in crops in China and Southeast Asia [6,7].…”

Section: Introductionmentioning

confidence: 99%

Analysis and application of the SRBSDV P9-1 octamer crystal structure for the target of α-amino phosphonate derivatives

Ding¹,

Li²,

Chen³

et al. 2018

Oncotarget

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Southern rice black-streaked dwarf virus (SRBSDV) P9-1 octameric protein accumulates viroplasms in SRBSDV-infected plant and insect cells, our previous studies found α-amino phosphonate drug-dufulin had a micromole affinity with SRBSDV P9-1. Now we focus our studies on the SRBSDV P9-1 crystal structure and use it as the target for α-amino phosphonate derivatives. The structure of the SRBSDV P9-1 cylindrical octamer was determined to a 2.2 Å resolution using X-ray crystallography, this structure was composed of nine α-helices, nine β-sheets and a series of interconnecting loops. The structures of all eight subunits were nearly identical, except for some differences in the β-sheet core. There were six different sites (R20K, V109D, F164T, V123L, C124L and V128T) present between the SRBSDV P9-1 and the previously reported RBSDV P9-1 crystal structure. Fluorescence titration, isothermal calorimetry and microscale thermophoresis experiments showed that α-amino phosphonate derivatives GUFCC-013 and GUFCC-023 bound to SRBSDV P9-1 with micromole binding affinities. These results will provide important help for the further improvement of the lead compound and develop the potential anti-SRBSDV drugs.

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“…Continuous cultures of cells derived from insect vectors are powerful tools for studying the replication cycle of persistent propagative plant viruses because they are more uniform than primary cultures and thus provide more consistent data (Creamer, 1993). Continuous cultures of leafhopper cells provide a good system to study the replication cycle of phytoreoviruses in their insect vectors (Kimura & Omura, 1988;Omura & Kimura, 1994;Wei et al, 2006Wei et al, , 2007.Rice stripe virus (RSV), the type species of the genus Tenuivirus, has filamentous ribonucleoprotein particles (RNPs) that contain nucleoprotein, RNA-dependent RNA polymerase and four negative-sense single-stranded RNA segments that encode seven proteins (Toriyama, 1986;Falk & Tsai, 1998;Wei et al, 2009;Xiao et al, 2010). RSV is transmitted by the small brown planthopper (SBPH; Laodelphax striatellus Fallén) in a persistent propagative manner (Toriyama, 1986;Wei et al, 2009).…”

mentioning

confidence: 99%

An insect cell line derived from the small brown planthopper supports replication of rice stripe virus, a tenuivirus

Chen

et al. 2013

Journal of General Virology

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A cell line from the small brown planthopper (SBPH; Laodelphax striatellus) was established to study replication of rice stripe virus (RSV), a tenuivirus. The SBPH cell line, which had been subcultured through 30 passages, formed monolayers of epithelial-like cells. Inoculation of cultured vector cells with RSV resulted in a persistent infection. During viral infection in the SBPH cell line, the viral non-structural protein NS3 co-localized with the filamentous ribonucleoprotein particles of RSV, as revealed by electron and confocal microscopy. The knockdown of NS3 expression due to RNA interference induced by synthesized double-stranded RNAs from the NS3 gene significantly inhibited viral infection in the SBPH cell line. These results demonstrated that NS3 of RSV might be involved in viral replication or assembly. The persistent infection of the SBPH cell line by RSV will enable a better understanding of the complex relationship between RSV and its insect vector.Plant viruses in the genera Tospovirus, Tenuivirus, Rhabdovirus, Phytoreovirus and Fijivirus are transmitted by their respective insect vectors in a persistent propagative manner (Ammar et al., 2009;Hogenhout et al., 2008). Insights into replication of these viruses in their vectors will help in development of new strategies to control the transmission of the viruses by the vectors. Continuous cultures of cells derived from insect vectors are powerful tools for studying the replication cycle of persistent propagative plant viruses because they are more uniform than primary cultures and thus provide more consistent data (Creamer, 1993). Continuous cultures of leafhopper cells provide a good system to study the replication cycle of phytoreoviruses in their insect vectors (Kimura & Omura, 1988;Omura & Kimura, 1994;Wei et al., 2006Wei et al., , 2007.Rice stripe virus (RSV), the type species of the genus Tenuivirus, has filamentous ribonucleoprotein particles (RNPs) that contain nucleoprotein, RNA-dependent RNA polymerase and four negative-sense single-stranded RNA segments that encode seven proteins (Toriyama, 1986;Falk & Tsai, 1998;Wei et al., 2009;Xiao et al., 2010). RSV is transmitted by the small brown planthopper (SBPH; Laodelphax striatellus Fallén) in a persistent propagative manner (Toriyama, 1986;Wei et al., 2009). Currently, the mechanism that enables efficient multiplication of RSV in its vector is unknown due to the lack of SBPH cell lines. In previous attempts to establish primary cell cultures, only a small number of cells migrated occasionally from the embryonic explants dissected from SBPH eggs, and eventually the subculture died off (Mitsuhashi, 1969(Mitsuhashi, , 2001Yamada et al., 1970). The present report describes the first successful establishment of continuous cultures of SBPHs for the study of RSV replication.The SBPH cell line was established by adapting the protocol described by Kimura & Omura (1988). Nonviruliferous SBPHs eggs that had been oviposited 8 days earlier in leaf sheaths of rice plants were surface-sterilized with 70 % ...

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The Spread of Rice Dwarf Virus among Cells of Its Insect Vector Exploits Virus-Induced Tubular Structures

Cited by 93 publications

References 50 publications

The P2 capsid protein of the nonenveloped rice dwarf phytoreovirus induces membrane fusion in insect host cells

The P2 capsid protein of the nonenveloped rice dwarf phytoreovirus induces membrane fusion in insect host cells

Analysis and application of the SRBSDV P9-1 octamer crystal structure for the target of α-amino phosphonate derivatives

An insect cell line derived from the small brown planthopper supports replication of rice stripe virus, a tenuivirus

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