Aquareovirus protein VP6 colocalizes with NS80 protein in infected and transfected cells

Wen, Dawei; Yan, Liming; Shao, Lingyun; Guo, Hongbo; Li, Xiaoming; Qin, Fang

doi:10.1186/1743-422x-10-133

Cited by 7 publications

(10 citation statements)

References 29 publications

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“…The VP2, VP4, VP5, VP6, VP7 and NS80 polyclonal antibodies (pAbs) of GCRV were prepared and stored in our laboratory [ 40 , 41 , 47 , 48 ]. VP1 and VP3 conserved regions were cloned into pRSET vector respectively based on GenBank sequences AF260511 and AF260513, and mouse polyclonal sera were prepared as described previously [ 48 ].…”

Section: Methodsmentioning

confidence: 99%

“…Plasmids pCI-neo-VP4, pCI-neo-VP6 and pCI-neo-NS80 were prepared previously [ 47 , 48 ]. The VP1, VP2, VP3, VP5 or VP7 gene amplified from cDNA of GCRV based on GenBank sequence (AF260511, AF260512, AF260513, AF403392, AF403396), was cloned into pCI-neo vector (Promega, USA) and named pCI-neo-VP1, pCI-neo-VP2, pCI-neo-VP3, pCI-neo-VP5 and pCI-neo-VP7 respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Immunofluorescence (IF) assays were performed as described previously [ 47 ]. Briefly, Vero cells were transfected with indicated plasmids according to the user manual of Lipofectamine 2000 (Lipo 2000) (Invitrogen, Carlsbad, USA) and fixed at 24 h post-transfection.…”

Section: Methodsmentioning

confidence: 99%

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Aquareovirus NS80 Initiates Efficient Viral Replication by Retaining Core Proteins within Replication-Associated Viral Inclusion Bodies

et al. 2015

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Viral inclusion bodies (VIBs) are specific intracellular compartments for reoviruses replication and assembly. Aquareovirus nonstructural protein NS80 has been identified to be the major constituent for forming globular VIBs in our previous study. In this study, we investigated the role of NS80 in viral structural proteins expression and viral replication. Immunofluorescence assays showed that NS80 could retain five core proteins or inner-capsid proteins (VP1-VP4 and VP6), but not outer-capsid proteins (VP5 and VP7), within VIBs in co-transfected or infected cells. Further co-immunoprecipitation analysis confirmed that NS80 could interact with each core protein respectively. In addition, we found that newly synthesized viral RNAs co-localized with VIBs. Furthermore, time-course analysis of viral structural proteins expression showed that the expression of NS80 was detected first, followed by the detection of inner shell protein VP3, and then of other inner-capsid proteins, suggesting that VIBs were essential for the formation of viral core frame or progeny virion. Moreover, knockdown of NS80 by shRNA not only inhibited the expression of aquareovirus structural proteins, but also inhibited viral infection. These results indicated that NS80-based VIBs were formed at earlier stage of infection, and NS80 was able to coordinate the expression of viral structural proteins and viral replication.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Aquareovirus NS80 Initiates Efficient Viral Replication by Retaining Core Proteins within Replication-Associated Viral Inclusion Bodies

et al. 2015

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“…GCRV was first isolated in 1983 and is characterized by severe hemorrhagic disease, with approximately 85% mortality in fingerling and yearling grass carp (Cyenopharyngodon idellus) in China. The GCRV is classified to species Aquareovirus-C in the genus Aquareovirus, which was thought to be most pathogenic aquareovirus among all the isolates reported to date (Wen et al 2013). Despite extensive investigation in the past 20 years, the pathogenic mechanisms by which attenuated strains and virulent strain can lead to hemorrhagic death in grass carp have not been well characterized, with a lack of information pertaining to GCRV tissue distributions a potentially important piece.…”

Section: Discussionmentioning

confidence: 99%

The distribution of different virulence grass carp reovirus strains in some neglected tissues

Liang¹,

Fu²,

Li³

et al. 2016

Polish Journal of Veterinary Sciences

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Grass carp reovirus (GCRV) is the causative agent of hemorrhagic disease in infected grass carp. During an outbreak, a mortality rate of up to 85% can be experienced, thus leading to substantial economic losses.The current understanding of disease pathogenesis is limited, with the distribution and dynamics of replication amongst different GCRV strains in vivo largely unknown. We determined distribution of different GCRV strains in infected grass carp, especially in some neglected tissues, such as the gill, brain, blood and so on. The results showed elevated viral RNA copy numbers in the blood, with some tissues such as the kidney, heart, brain, and bladder exhibiting even higher viral loads following infection with the virulent GCRV-CL strain. Even more interesting is that the brain exhibited the highest viral load, with a copy number of 800,000 following GCRV-CL infection. Overall, this study provides further insight into GCRV viral load distributions following infection and potentially identified some new viral tropism sites to provide a foundation for further studies aimed at characterizing GCRV viral pathogenesis.

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“…r2 is also a potent dsRNA-binding protein that binds to the base of l1 for outer capsid assembly [18]. Therefore, It has also been hypothesized that GCRV-AH528 VP6 functions as a mediator bridging the inner core with the outer capsid [32].…”

Section: Discussionmentioning

confidence: 99%

Structure and function of S9 segment of grass carp reovirus Anhui strain

Jiang

et al. 2017

VirusDis.

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A highly virulent grass carp reovirus (GCRV) strain, named GCRV-AH528, was recently purified from a diseased grass carp with hemorrhage disease in Anhui, China. GCRV-AH528 S9 segment was 1320 nucleotides in length and encoded a 418 amino acid VP6 protein. BLAST search showed that the VP6 protein owned a conserved domain belonging to the reoviral r2 family. Phylogenetic analysis of VP6 presented that GCRV-AH528 belonged to GCRV genotype II, which was more closely related to Orthoreovirus than GCRV genotype I and genotype III. Further analysis revealed that GCRV-AH528 S9 and mammalian orthoreovirus S8 might have evolved from a common ancestral precursor and have identical mechanism in virus assembly. The expression level of vp6 gene was detected by quantitative real-time PCR (qRT-PCR). Over time, the expression level of vp6 gradually increased in Ctenopharyngodon idellus kidney cells. However, the level of vp6 expression in blood sharply increased at 4-6 days, and then decreased to a low level after GCRV-AH528 challenge (P \ 0.05). The vp6 gene was detected in all tissues examined, whereas at relatively higher levels in blood, kidney, and liver (P \ 0.05). The yeast two-hybrid (Y2H) system was used to identify VP6 self-interaction, while no interaction was detected in VP6-VP6. This study not only revealed the S9 segment structure and expression pattern but also analyzed the VP6 mechanism by yeast hybridization method. The present study provides valuable informations for further experimental design and investigation of VP6 functions.

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Aquareovirus protein VP6 colocalizes with NS80 protein in infected and transfected cells

Cited by 7 publications

References 29 publications

Aquareovirus NS80 Initiates Efficient Viral Replication by Retaining Core Proteins within Replication-Associated Viral Inclusion Bodies

Aquareovirus NS80 Initiates Efficient Viral Replication by Retaining Core Proteins within Replication-Associated Viral Inclusion Bodies

The distribution of different virulence grass carp reovirus strains in some neglected tissues

Structure and function of S9 segment of grass carp reovirus Anhui strain

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