The Experts below are selected from a list of 27 Experts worldwide ranked by ideXlab platform
Liqun Lu - One of the best experts on this subject based on the ideXlab platform.
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Identification of a novel membrane-associated protein from the S7 segment of grass carp reovirus
Journal of General Virology, 2019Co-Authors: Fei Yu, Longlong Wang, Wanjuan Li, Liqun LuAbstract:Aquareovirus is a genus of viruses in family Reoviridae, subfamily Spinareovirinae, members of which infect fish, shellfish and crustaceans. Grass carp reovirus (GCRV), a genotype 1 reovirus isolated from grass carp, has served as a model strain for investigating aquareovirus–host interactions. Herein, we report a neglected open reading frame (ORF), tentatively named NS12, residing between NS16 and NS31 in segment 7 (S7) of the virus genome. With an additional reading frame, the nucleotide sequence of NS12 partially overlaps with the 3′ expressible nucleotide sequence of NS16. NS12 is not a pseudogene during virus replication, as confirmed in fish cells infected with GCRV and based on amino acid sequence analysis and protein expression pattern. Bioinformatics analysis indicated that NS12 is a transmembrane protein, which was confirmed by its exclusive presence in the membrane-associated fraction of the cell lysate. However, unlike fusion protein NS16, NS12 alone could not induce visible syncytium formation in fish cells. Thus, NS12 is functionally distinct from known aquareovirus membrane-associated protein NS16. NS12-like ORFs (with an AUG or non-AUG initiator codon) are also present in the S7 segment of other aquareoviruses, suggesting that NS12 homologues may be widely distributed in the genus Aquareovirus.
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Characterization of the interaction between outer-fiber protein VP55 of genotype III grass carp reovirus and Fibulin-4 of grass carp.
Fish & Shellfish Immunology, 2018Co-Authors: Longlong Wang, Fei Yu, Liqun LuAbstract:Abstract Genotype III grass carp reovirus (GCRV; representative strain, GCRV-104) belongs to the subfamily Spinareovirinae and encodes an outer-fiber protein, VP55, responsible for mediating the infection of target tissues by the virus and assisting the virus into cells. Fibulin-4/EFEMP2 protein was previously identified as a putative binding partner for VP55 in a yeast two-hybrid screening. Here, we have further characterized the association between Fibulin-4 and VP55 by using protein interaction assays. An intracellular co-localization assay showed that RFP-Fibulin-4 co-localized with GFP-VP55 in grass carp ovary (GCO) cells. Bacterially expressed GST-tagged Fibulin-4 was shown to associate with baculovirus-expressed His-tagged VP55 in a dot-blot overlay assay; moreover, baculovirus-expressed His-tagged VP55 was able to pull down GFP-Fibulin-4 expressed in the GCO cells. We performed real-time PCR and immunoblotting analysis and showed that endogenous Fibulin-4, although suppressed to a lower level in the late infection phase, is present throughout the infection course of GCRV-104 in CIK cells. In conclusion, our results indicate that grass carp Fibulin-4 interacts with VP55. The presence of Fibulin-4, a well-known secreted protein, during the infection course of GCRV-104 in grass carp cells implies its potential role during viral egression through interaction with VP55.
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Inhibitor analysis revealed that clathrin-mediated endocytosis is involed in cellular entry of type III grass carp reovirus.
Virology Journal, 2018Co-Authors: Hao Wang, Dubo Chen, Lingbing Zeng, Liqun LuAbstract:Grass carp (Ctenopharyngodon idella) hemorrhagic disease is caused by an acute infection with grass carp reovirus (GCRV). The frequent outbreaks of this disease have suppressed development of the grass carp farming industry. GCRV104, the representative strain of genotype III grass carp (Ctenopharyngodon idella) reovirus, belongs to the Spinareovirinae subfamily and serves as a model for studying the strain of GCRV which encodes an outer-fiber protein. There is no commercially available vaccine for this genotype of GCRV. Therefore, the discovery of new inhibitors for genotype III of GCRV will be clinically beneficial. In addition, the mechanism of GCRV with fiber entry into cells remains poorly understood. Viral entry was determined by a combination of specific pharmacological inhibitors, transmission electron microscopy, and real-time quantitative PCR. Our results demonstrate that both GCRV-JX01 (genotype I) and GCRV104 (genotype III) of GCRV propagated in the grass carp kidney cell line (CIK) with a typical cytopathic effect (CPE). However, GCRV104 replicated slower than GCRV-JX01 in CIK cells. The titer of GCRV-JX01 was 1000 times higher than GCRV104 at 24 h post-infection. We reveal that ammonium chloride, dynasore, pistop2, chlorpromazine, and rottlerin inhibit viral entrance and infection, but not nystatin, methyl-β-cyclodextrin, IPA-3, amiloride, bafilomycin A1, nocodazole, and latrunculin B. Furthermore, GCRV104 and GCRV-JX01 infection of CIK cells depended on dynamin and the acidification of the endosome. This was evident by the significant inhibition following prophylactic treatment with the lysosomotropic drug ammonium chloride or dynasore. Taken together, our data have suggested that GCRV104 enters CIK cells through clathrin-mediated endocytosis in a pH-dependent manner. We also suggest that dynamin is critical for efficient viral entry. Additionally, the phosphatidylinositol 3-kinase inhibitor wortmannin and the protein kinase C inhibitor rottlerin block GCRV104 cell entry and replication.
Longlong Wang - One of the best experts on this subject based on the ideXlab platform.
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Identification of a novel membrane-associated protein from the S7 segment of grass carp reovirus
Journal of General Virology, 2019Co-Authors: Fei Yu, Longlong Wang, Wanjuan Li, Liqun LuAbstract:Aquareovirus is a genus of viruses in family Reoviridae, subfamily Spinareovirinae, members of which infect fish, shellfish and crustaceans. Grass carp reovirus (GCRV), a genotype 1 reovirus isolated from grass carp, has served as a model strain for investigating aquareovirus–host interactions. Herein, we report a neglected open reading frame (ORF), tentatively named NS12, residing between NS16 and NS31 in segment 7 (S7) of the virus genome. With an additional reading frame, the nucleotide sequence of NS12 partially overlaps with the 3′ expressible nucleotide sequence of NS16. NS12 is not a pseudogene during virus replication, as confirmed in fish cells infected with GCRV and based on amino acid sequence analysis and protein expression pattern. Bioinformatics analysis indicated that NS12 is a transmembrane protein, which was confirmed by its exclusive presence in the membrane-associated fraction of the cell lysate. However, unlike fusion protein NS16, NS12 alone could not induce visible syncytium formation in fish cells. Thus, NS12 is functionally distinct from known aquareovirus membrane-associated protein NS16. NS12-like ORFs (with an AUG or non-AUG initiator codon) are also present in the S7 segment of other aquareoviruses, suggesting that NS12 homologues may be widely distributed in the genus Aquareovirus.
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Characterization of the interaction between outer-fiber protein VP55 of genotype III grass carp reovirus and Fibulin-4 of grass carp.
Fish & Shellfish Immunology, 2018Co-Authors: Longlong Wang, Fei Yu, Liqun LuAbstract:Abstract Genotype III grass carp reovirus (GCRV; representative strain, GCRV-104) belongs to the subfamily Spinareovirinae and encodes an outer-fiber protein, VP55, responsible for mediating the infection of target tissues by the virus and assisting the virus into cells. Fibulin-4/EFEMP2 protein was previously identified as a putative binding partner for VP55 in a yeast two-hybrid screening. Here, we have further characterized the association between Fibulin-4 and VP55 by using protein interaction assays. An intracellular co-localization assay showed that RFP-Fibulin-4 co-localized with GFP-VP55 in grass carp ovary (GCO) cells. Bacterially expressed GST-tagged Fibulin-4 was shown to associate with baculovirus-expressed His-tagged VP55 in a dot-blot overlay assay; moreover, baculovirus-expressed His-tagged VP55 was able to pull down GFP-Fibulin-4 expressed in the GCO cells. We performed real-time PCR and immunoblotting analysis and showed that endogenous Fibulin-4, although suppressed to a lower level in the late infection phase, is present throughout the infection course of GCRV-104 in CIK cells. In conclusion, our results indicate that grass carp Fibulin-4 interacts with VP55. The presence of Fibulin-4, a well-known secreted protein, during the infection course of GCRV-104 in grass carp cells implies its potential role during viral egression through interaction with VP55.
Fei Yu - One of the best experts on this subject based on the ideXlab platform.
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Identification of a novel membrane-associated protein from the S7 segment of grass carp reovirus
Journal of General Virology, 2019Co-Authors: Fei Yu, Longlong Wang, Wanjuan Li, Liqun LuAbstract:Aquareovirus is a genus of viruses in family Reoviridae, subfamily Spinareovirinae, members of which infect fish, shellfish and crustaceans. Grass carp reovirus (GCRV), a genotype 1 reovirus isolated from grass carp, has served as a model strain for investigating aquareovirus–host interactions. Herein, we report a neglected open reading frame (ORF), tentatively named NS12, residing between NS16 and NS31 in segment 7 (S7) of the virus genome. With an additional reading frame, the nucleotide sequence of NS12 partially overlaps with the 3′ expressible nucleotide sequence of NS16. NS12 is not a pseudogene during virus replication, as confirmed in fish cells infected with GCRV and based on amino acid sequence analysis and protein expression pattern. Bioinformatics analysis indicated that NS12 is a transmembrane protein, which was confirmed by its exclusive presence in the membrane-associated fraction of the cell lysate. However, unlike fusion protein NS16, NS12 alone could not induce visible syncytium formation in fish cells. Thus, NS12 is functionally distinct from known aquareovirus membrane-associated protein NS16. NS12-like ORFs (with an AUG or non-AUG initiator codon) are also present in the S7 segment of other aquareoviruses, suggesting that NS12 homologues may be widely distributed in the genus Aquareovirus.
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Characterization of the interaction between outer-fiber protein VP55 of genotype III grass carp reovirus and Fibulin-4 of grass carp.
Fish & Shellfish Immunology, 2018Co-Authors: Longlong Wang, Fei Yu, Liqun LuAbstract:Abstract Genotype III grass carp reovirus (GCRV; representative strain, GCRV-104) belongs to the subfamily Spinareovirinae and encodes an outer-fiber protein, VP55, responsible for mediating the infection of target tissues by the virus and assisting the virus into cells. Fibulin-4/EFEMP2 protein was previously identified as a putative binding partner for VP55 in a yeast two-hybrid screening. Here, we have further characterized the association between Fibulin-4 and VP55 by using protein interaction assays. An intracellular co-localization assay showed that RFP-Fibulin-4 co-localized with GFP-VP55 in grass carp ovary (GCO) cells. Bacterially expressed GST-tagged Fibulin-4 was shown to associate with baculovirus-expressed His-tagged VP55 in a dot-blot overlay assay; moreover, baculovirus-expressed His-tagged VP55 was able to pull down GFP-Fibulin-4 expressed in the GCO cells. We performed real-time PCR and immunoblotting analysis and showed that endogenous Fibulin-4, although suppressed to a lower level in the late infection phase, is present throughout the infection course of GCRV-104 in CIK cells. In conclusion, our results indicate that grass carp Fibulin-4 interacts with VP55. The presence of Fibulin-4, a well-known secreted protein, during the infection course of GCRV-104 in grass carp cells implies its potential role during viral egression through interaction with VP55.
Drake C. Stenger - One of the best experts on this subject based on the ideXlab platform.
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Reovirus genomes from plant-feeding insects represent a newly discovered lineage within the family Reoviridae
Virus Research, 2011Co-Authors: Allyn Spear, Mark S. Sisterson, Drake C. StengerAbstract:Abstract A complex set of double-stranded RNAs (dsRNAs) was isolated from threecornered alfalfa hopper (Spissistilus festinus), a plant-feeding hemipteran pest. A subset of these dsRNAs constitute the genome of a new reovirus, provisionally designated Spissistilus festinus reovirus (SpFRV). SpFRV was present in threecornered alfalfa hopper populations in the San Joaquin Valley of California, with incidence ranging from 10% to 60% in 24 of 25 sample sets analyzed. The 10 dsRNA segments of SpFRV were completely sequenced and shown to share conserved terminal sequences (5′-AGAGA and CGAUGUUGU-3′) of the positive-sense strand that are distinct from known species of the family Reoviridae. Comparisons of the RNA directed RNA polymerase (RdRp) indicated SpFRV is most closely related (39.1% amino acid identity) to another new reovirus infecting the angulate leafhopper (Acinopterus angulatus) and provisionally designated Acinopterus angulatus reovirus (AcARV). The RdRp of both viruses was distantly related to Raspberry latent virus RdRp at 27.0% (SpFRV) and 30.0% (AcARV) or Rice ragged stunt virus RdRp at 26.2% (SpFRV) and 29.0% (AcARV) amino acid identity. RdRp phylogeny confirmed that SpFRV and AcARV are sister taxa sharing a most recent common ancestor. SpFRV segment 6 encodes a protein containing two NTP binding motifs that are conserved in homologs of reoviruses in the subfamily Spinareovirinae. The protein encoded by SpFRV segment 4 was identified as a guanylyltransferase homolog. SpFRV segments 1, 3, and 10 encode homologs of reovirus structural proteins. No homologs were identified for proteins encoded by SpFRV segments 5, 7, 8, and 9. Collectively, the low level of sequence identity with other reoviruses, similar segment terminal sequences, RdRp phylogeny, and host taxa indicate that SpFRV and AcARV may be considered members of a proposed new genus of the family Reoviridae (subfamily Spinareovirinae), with SpFRV assigned as the type species.
Tony L. Goldberg - One of the best experts on this subject based on the ideXlab platform.
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Pteropine Orthoreovirus in an Angolan Soft-Furred Fruit Bat (Lissonycteris angolensis) in Uganda Dramatically Expands the Global Distribution of an Emerging Bat-Borne Respiratory Virus.
Viruses, 2020Co-Authors: Andrew J. Bennett, Tony L. GoldbergAbstract:Pteropine orthoreovirus (PRV; Reoviridae: Spinareovirinae) is an emerging bat-borne zoonotic virus that causes influenza-like illness (ILI). PRV has thus far been found only in Australia and Asia, where diverse old-world fruit bats (Pteropodidae) serve as hosts. In this study, we report the discovery of PRV in Africa, in an Angolan soft-furred fruit bat (Lissonycteris angolensis ruwenzorii) from Bundibugyo District, Uganda. Metagenomic characterization of a rectal swab yielded 10 dsRNA genome segments, revealing this virus to cluster within the known diversity of PRV variants detected in bats and humans in Southeast Asia. Phylogeographic analyses revealed a correlation between geographic distance and genetic divergence of PRVs globally, which suggests a geographic continuum of PRV diversity spanning Southeast Asia to sub-Saharan Africa. The discovery of PRV in an African bat dramatically expands the geographic range of this zoonotic virus and warrants further surveillance for PRVs outside of Southeast Asia.