The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Michael M. Goodin - One of the best experts on this subject based on the ideXlab platform.
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Lost and found: Rediscovery and genomic characterization of sowthistle yellow vein virus after a 30+ year hiatus.
Virus Research, 2020Co-Authors: Drake C. Stenger, Lindsey P. Burbank, Renyuan Wang, Alexander A. Stewart, Caleb Mathias, Michael M. GoodinAbstract:Abstract Beginning in the 1960’s, sowthistle yellow vein virus (SYVV) was the subject of pioneering research that demonstrated propagation of a plant virus in its insect vector. Since the 1980’s there has been a paucity of research on SYVV, with historic isolates no longer maintained and no genomic sequence available. Once commonly observed infecting sowthistle (Sonchus oleraceous L.) in California, SYVV incidence declined ca. 1990, likely due to displacement of the black currant aphid (Hyperomyzus lactucae L.) by an invasive non-vector aphid. In 2018, SYVV was fortuitously rediscovered infecting sowthistle in an organic citrus grove in Kern County, CA. The SYVV genome sequence (13,719 nts) obtained from the 2018 sample (designated HWY65) encoded all six expected genes: N, P, MP, M, G, and L. Nucleotide sequence (representing ∼86 % of the genome) of the SYVV Berkeley lab isolate, used by E. S. Sylvester and colleagues for the paradigm-shifting research mentioned above, was determined from an archived library of cDNA clones constructed in 1986. The two nucleotide sequences share 98.5 % identity, confirming both represent the same virus, thereby linking biology of the historic isolate with extant SYVV rediscovered in 2018. Phylogenetic analysis of the L protein indicated SYVV is positioned within a clade containing a subset of viruses currently assigned to the genus Nucleorhabdovirus. As Nucleorhabdovirus is paraphyletic, the International Committee on the Taxonomy of Viruses has proposed abolishment of the genus and establishment of three new genera. In this revised taxonomy, the clade containing SYVV constitutes a new genus designated BetaNucleorhabdovirus.
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Sonchus yellow net virus core particles form on ring-like nuclear structure enriched in viral phosphoprotein.
Virus Research, 2018Co-Authors: Tea Meulia, Lucy R. Stewart, Michael M. GoodinAbstract:The phosphoprotein (P) of the Nucleorhabdovirus sonchus yellow net virus has been shown to accumulate in ring-shaped structures in virus-infected nuclei. Further examination by live-cell imaging, in combination with structural examination by transmission electron microscopy and immunolocalization demonstrated that P-rings do not form in association with nucleoli. Furthermore, viral cores were shown to condense on the nucleoplasm-contacting surface of the rings. The data presented here offer evidence for the site of nucleocapsid assembly in SYNV-infected nuclei.
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Subcellular Localization and Nuclear Import of Maize Fine Streak Virus and Maize Mosaic Virus Proteins
2015Co-Authors: Chi-wei Tsai, Valdir R. Correa, Margaret G. Redinbaugh, Michael M. Goodin, Saskia A. HogenhoutAbstract:Maize fine streak virus (MFSV) and Maize mosaic virus (MMV) are members of the genus Nucleorhabdovirus in the family Rhabdoviridae. Plant rhabdoviruses are divided into two genera, Nucleorhabdovirus and Cytorhabdovirus. Nucleorhabdoviruses assemble at inner nuclear envelopes, whereas cytorhabdoviruses assemble at cytoplasmic membranes. The MFSV genome encodes seven proteins in the gene order 3’-N-P-3-4-M-G-L-5’, and the MMV genome encodes six proteins in the order 3’-N-P-3-M-G-L-5 ’ (Fig. 1). Nucleorhabdoviruses assemble in the nuclei of their plant and insect hosts, and therefore nuclear import of viral proteins is critical to complete morphogenesis. Nuclear import of nuclear localization signal (NLS)-containing proteins is mediated by Importin a and b. Importin a binds NLS-containing protein, and this heterodimer subsequently binds Importin b. The tripartite complex then docks to the nuclear pore followed by translocation into the nucleus. We show that the MFSV N and P complex colocalizes to the nucleolus and is dependent on Importin a for nuclear import. This is the first demonstration that Importin a is involved in nuclear import of rhabdoviral proteins in plant cells
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The Nucleocapsid Protein of Potato Yellow dwarf Virus: Protein Interactions and Nuclear Import Mediated by a Non-Canonical Nuclear Localization Signal
Frontiers in Plant Science, 2012Co-Authors: Gavin Anderson, Anindya Bandyopadhyay, Renyuan Wang, Michael M. GoodinAbstract:Potato yellow dwarf virus (PYDV) is the type species of the genus Nucleorhabdovirus and, like all members of this genus, replication and morphogenesis occurs inside the nuclei of infected cells. Protein localization prediction algorithms failed to identify a nuclear localization signal (NLS) in PYDV nucleocapsid (N) protein, although PYDV-N has been shown to localize exclusively to the nucleus when expressed as a GFP:N fusion in plant cells. Deletion analysis using fragments of PYDV-N identified a karyophillic region in the carboxy-terminal 122 amino acids. Alanine-scanning mutagenesis was performed across this region in the context of the full-length N protein. Mutants were assayed for their ability to nuclear localize using live-cell imaging and a yeast-based assay. Two amino acid motifs, 419QKR421 and 432KR433 were shown to be essential for nuclear import and interaction with importin-α. Additional bimolecular fluorescence complementation (BiFC) showed that the PYDV-N NLS mutants cannot be ferried into the nucleus via interaction with PYDV-P or -M. In contrast, interaction with N NLS mutants appeared to retard the nuclear import of PYDV-P. GFP fused to aa 419-434 established that the PYDV-N-NLS can function outside the context of this protein. Taken together, it was determined that PYDV-N contains the bipartite NLS 419QKRANEEAPPAAQKR433.
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An integrated protein localization and interaction map for Potato yellow dwarf virus, type species of the genus Nucleorhabdovirus.
Virology, 2010Co-Authors: Anindya Bandyopadhyay, Kathleen M. Martin, Kristin Kopperud, Gavin Anderson, Michael M. GoodinAbstract:The genome of Potato yellow dwarf virus (PYDV; Nucleorhabdovirus type species) was determined to be 12,875 nucleotides (nt). The antigenome is organized into seven open reading frames (ORFs) ordered 3'-N-X-P-Y-M-G-L-5', which likely encode the nucleocapsid, phospho, movement, matrix, glyco and RNA-dependent RNA polymerase proteins, respectively, except for X, which is of unknown function. The ORFs are flanked by a 3' leader RNA of 149 nt and a 5' trailer RNA of 97 nt, and are separated by conserved intergenic junctions. Phylogenetic analyses indicated that PYDV is closely related to other leafhopper-transmitted rhabdoviruses. Functional protein assays were used to determine the subcellular localization of PYDV proteins. Surprisingly, the M protein was able to induce the intranuclear accumulation of the inner nuclear membrane in the absence of any other viral protein. Finally, bimolecular fluorescence complementation was used to generate the most comprehensive protein interaction map for a plant-adapted rhabdovirus to date.
Nicolás Bejerman - One of the best experts on this subject based on the ideXlab platform.
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Novel bird’s-foot trefoil RNA viruses provide insights into a clade of legume-associated enamoviruses and rhabdoviruses
Archives of Virology, 2019Co-Authors: Humberto J. Debat, Nicolás BejermanAbstract:Here, we report the identification and characterization of two novel viruses associated with bird’s-foot trefoil. Virus sequences related to those of enamoviruses (ssRNA (+); Luteoviridae; Enamovirus ) and Nucleorhabdoviruses (ssRNA (-); Rhabdoviridae; Nucleorhabdovirus ) were detected in Lotus corniculatus transcriptome data. The genome of the tentatively named “bird’s-foot trefoil-associated virus 1” (BFTV-1) is a 13,626-nt-long negative-sense ssRNA. BFTV-1 encodes six predicted gene products in the antigenome orientation in the canonical order 3′-N-P-P3-M-G-L-5′. The genome of the proposed “bird’s-foot trefoil-associated virus 2” (BFTV-2) is 5,736 nt long with a typical 5΄-PO-P1-2-IGS-P3-P5-3′ enamovirus genome structure. Phylogenetic analysis indicated that BFTV-1 is closely related to datura yellow vein Nucleorhabdovirus and that BFTV-2 clusters into a monophyletic lineage of legume-associated enamoviruses. This subclade of highly related and co-divergent legume-associated viruses provides insights into the evolutionary history of the enamoviruses.
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Novel bird's-foot trefoil RNA viruses provide insights into a clade of legume-associated enamoviruses and rhabdoviruses.
Archives of Virology, 2019Co-Authors: Humberto J. Debat, Nicolás BejermanAbstract:Here, we report the identification and characterization of two novel viruses associated with bird’s-foot trefoil. Virus sequences related to those of enamoviruses (ssRNA (+); Luteoviridae; Enamovirus) and Nucleorhabdoviruses (ssRNA (-); Rhabdoviridae; Nucleorhabdovirus) were detected in Lotus corniculatus transcriptome data. The genome of the tentatively named “bird’s-foot trefoil-associated virus 1” (BFTV-1) is a 13,626-nt-long negative-sense ssRNA. BFTV-1 encodes six predicted gene products in the antigenome orientation in the canonical order 3′-N-P-P3-M-G-L-5′. The genome of the proposed “bird’s-foot trefoil-associated virus 2” (BFTV-2) is 5,736 nt long with a typical 5΄-PO-P1-2-IGS-P3-P5-3′ enamovirus genome structure. Phylogenetic analysis indicated that BFTV-1 is closely related to datura yellow vein Nucleorhabdovirus and that BFTV-2 clusters into a monophyletic lineage of legume-associated enamoviruses. This subclade of highly related and co-divergent legume-associated viruses provides insights into the evolutionary history of the enamoviruses.
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Identification of novel RNA viruses associated to bird’s-foot trefoil (Lotus corniculatus)
2018Co-Authors: Humberto J. Debat, Nicolás BejermanAbstract:Bird9s-foot trefoil ( Lotus corniculatus L.) is a highly nutritious forage crop, employed for livestock foraging around the world. Despite the agronomical importance of this resilient crop, the related literature is rather scarce. Here, we report the identification and characterization of two novel viruses associated with bird9s-foot trefoil. Virus sequences with affinity to enamoviruses (ssRNA (+); Luteoviridae ; Enamovirus ) and Nucleorhabdoviruses (ssRNA (-); Rhabdoviridae ; Nucleorhabdovirus ) were detected in L. corniculatus transcriptome data. The proposed bird9s-foot trefoil enamovirus 1 (BFTEV-1) 5,736 nt virus sequence presents a typical 59-PO-P1-2-IGS-P3-P5-39; enamovirus genome structure. The tentatively named bird9s-foot trefoil Nucleorhabdovirus (BFTNRV) genome organization is characterized by 13,626 nt long negative-sense single-stranded RNA. BFTNRV presents in its antigenome orientation six predicted gene products in the canonical order 39-N-P-P3-M-G-L-59. Phylogenetic analysis suggests that BFTNRV is closely related to Datura yellow vein Nucleorhabdovirus , and that BFTEV-1 clusters into a monophyletic clade of legumes-associated enamoviruses. The bioinformatic reanalysis of SRA libraries deposited in the NCBI database constitutes an emerging approach to the discovery of novel plant viruses. The RNA viruses reported here provide a first glimpse of the virus landscape of this important crop. Future studies should assess the prevalence of BFTEV-1 and BFTNRV, and unravel whether the infection of these novel viruses is associated to specific symptoms.
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Cytorhabdovirus P3 genes encode 30K-like cell-to-cell movement proteins.
Virology, 2016Co-Authors: Krin S. Mann, Nicolás Bejerman, Karyn N. Johnson, Ralf G. DietzgenAbstract:Plant viruses encode movement proteins (MP) to facilitate cell-to-cell transport through plasmodesmata. In this study, using trans-complementation of a movement-defective turnip vein-clearing tobamovirus (TVCV) replicon, we show for the first time for cytorhabdoviruses (lettuce necrotic yellows virus (LNYV) and alfalfa dwarf virus (ADV)) that their P3 proteins function as MP similar to the TVCV P30 protein. All three MP localized to plasmodesmata when ectopically expressed. In addition, we show that these MP belong to the 30K superfamily since movement was inhibited by mutation of an aspartic acid residue in the critical 30K-specific LxD/N50-70G motif. We also report that Nicotiana benthamiana microtubule-associated VOZ1-like transcriptional activator interacts with LNYV P3 and TVCV P30 but not with ADV P3 or any of the MP point mutants. This host protein, which is known to interact with P3 of sonchus yellow net Nucleorhabdovirus, may be involved in aiding the cell-to-cell movement of LNYV and TVCV.
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Complete genome sequence and intracellular protein localization of Datura yellow vein Nucleorhabdovirus
Virus Research, 2015Co-Authors: Ralf G. Dietzgen, David J. Innes, Nicolás BejermanAbstract:A limited number of plant rhabdovirus genomes have been fully sequenced, making taxonomic classification, evolutionary analysis and molecular characterization of this virus group difficult. We have for the first time determined the complete genome sequence of 13,188 nucleotides of Datura yellow vein Nucleorhabdovirus (DYVV). DYVV genome organization resembles that of its closest relative, Sonchus yellow net virus (SYNV), with six ORFs in antigenomic orientation, separated by highly conserved intergenic regions and flanked by complementary 3′ leader and 5′ trailer sequences. As is typical for Nucleorhabdoviruses, all viral proteins, except the glycoprotein, which is targeted to the endoplasmic reticulum, are localized to the nucleus. Nucleocapsid (N) protein, matrix (M) protein and polymerase, as components of nuclear viroplasms during replication, have predicted strong canonical nuclear localization signals, and N and M proteins exclusively localize to the nucleus when transiently expressed as GFP fusions. As in all Nucleorhabdoviruses studied so far, N and phosphoprotein P interact when co-expressed, significantly increasing P nuclear localization in the presence of N protein. This research adds to the list of complete genomes of plant-infecting rhabdoviruses, provides molecular tools for further characterization and supports classification of DYVV as a Nucleorhabdovirus closely related to but with some distinct differences from SYNV.
Andrew O Jackson - One of the best experts on this subject based on the ideXlab platform.
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the matrix protein of a plant rhabdovirus mediates superinfection exclusion by inhibiting viral transcription
Journal of Virology, 2019Co-Authors: Xin Zhou, Andrew O Jackson, Zhenghe Li, Xueping ZhouAbstract:Superinfection exclusion (SIE) or cross-protection phenomena have been documented for plant viruses for nearly a century and are widespread among taxonomically diverse viruses, but little information is available about SIE of plant negative-strand RNA viruses. Here, we demonstrate that SIE by sonchus yellow net Nucleorhabdovirus virus (SYNV) is mediated by the viral matrix (M) protein, a multifunctional protein involved in transcription regulation, virion assembly, and virus budding. We show that fluorescent protein-tagged SYNV variants display mutual exclusion/cross-protection in Nicotiana benthamiana plants. Transient expression of the SYNV M protein, but not other viral proteins, interfered with SYNV local infections. In addition, SYNV M deletion mutants failed to exclude superinfection by wild-type SYNV. An SYNV minireplicon reporter gene expression assay showed that the M protein inhibited viral transcription. However, M protein mutants with weakened nuclear localization signals (NLS) and deficient nuclear interactions with the SYNV nucleocapsid protein were unable to suppress transcription. Moreover, SYNV with M NLS mutations exhibited compromised SIE against wild-type SYNV. From these data, we propose that M protein accumulating in nuclei with primary SYNV infections either coils or prevents uncoiling of nucleocapsids released by the superinfecting SYNV virions and suppresses transcription of superinfecting genomes, thereby preventing superinfection. Our model suggests that the rhabdovirus M protein regulates the transition from replication to virion assembly and renders the infected cells nonpermissive for secondary infections.IMPORTANCE Superinfection exclusion (SIE) is a widespread phenomenon in which an established virus infection prevents reinfection by closely related viruses. Understanding the mechanisms governing SIE will not only advance our basic knowledge of virus infection cycles but may also lead to improved design of antiviral measures. Despite the significance of SIE, our knowledge about viral SIE determinants and their modes of actions remain limited. In this study, we show that sonchus yellow net virus (SYNV) SIE is mediated by the viral matrix (M) protein. During primary infections, accumulation of M protein in infected nuclei results in coiling of genomic nucleocapsids and suppression of viral transcription. Consequently, nucleocapsids released by potential superinfectors are sequestered and are unable to initiate new infections. Our data suggest that SYNV SIE is caused by M protein-mediated transition from replication to virion assembly and that this process prevents secondary infections.
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Specificity of Plant Rhabdovirus Cell-to-Cell Movement
Journal of Virology, 2019Co-Authors: Xin Zhou, Xueping Zhou, Kai Sun, Wenye Lin, Shuo Wang, Andrew O JacksonAbstract:Positive-stranded RNA virus movement proteins (MPs) generally lack sequence-specific nucleic acid-binding activities and display cross-family movement complementarity with related and unrelated viruses. Negative-stranded RNA plant rhabdoviruses encode MPs with limited structural and functional relatedness with other plant virus counterparts, but the precise mechanisms of intercellular transport are obscure. In this study, we first analyzed the abilities of MPs encoded by five distinct rhabdoviruses to support cell-to-cell movement of two positive-stranded RNA viruses by using trans-complementation assays. Each of the five rhabdovirus MPs complemented the movement of MP-defective mutants of tomato mosaic virus and potato X virus. In contrast, movement of recombinant MP deletion mutants of sonchus yellow net Nucleorhabdovirus (SYNV) and tomato yellow mottle-associated cytorhabdovirus (TYMaV) was rescued only by their corresponding MPs, i.e., SYNV sc4 and TYMaV P3. Subcellular fractionation analyses revealed that SYNV sc4 and TYMaV P3 were peripherally associated with cell membranes. A split-ubiquitin membrane yeast two-hybrid assay demonstrated specific interactions of the membrane-associated rhabdovirus MPs only with their cognate nucleoproteins (N) and phosphoproteins (P). More importantly, SYNV sc4-N and sc4-P interactions directed a proportion of the N-P complexes from nuclear sites of replication to punctate loci at the cell periphery that partially colocalized with the plasmodesmata. Our data show that cell-to-cell movement of plant rhabdoviruses is highly specific and suggest that cognate MP-nucleocapsid core protein interactions are required for intra- and intercellular trafficking. IMPORTANCE Local transport of plant rhabdoviruses likely involves the passage of viral nucleocapsids through MP-gated plasmodesmata, but the molecular mechanisms are not fully understood. We have conducted complementation assays with MPs encoded by five distinct rhabdoviruses to assess their movement specificity. Each of the rhabdovirus MPs complemented the movement of MP-defective mutants of two positive-stranded RNA viruses that have different movement strategies. In marked contrast, cell-to-cell movement of two recombinant plant rhabdoviruses was highly specific in requiring their cognate MPs. We have shown that these rhabdovirus MPs are localized to the cell periphery and associate with cellular membranes, and that they interact only with their cognate nucleocapsid core proteins. These interactions are able to redirect viral nucleocapsid core proteins from their sites of replication to the cell periphery. Our study provides a model for the specific inter- and intracellular trafficking of plant rhabdoviruses that may be applicable to other negative-stranded RNA viruses.
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Matrix-glycoprotein interactions required for budding of a plant Nucleorhabdovirus and induction of inner nuclear membrane invagination.
Molecular Plant Pathology, 2018Co-Authors: Kai Sun, Xin Zhou, Xueping Zhou, Wenye Lin, Andrew O JacksonAbstract:Nucleorhabdoviruses such as Sonchus yellow net virus (SYNV) replicate in the nuclei and undergo morphogenesis at the inner nuclear membrane (IM) in plant cells. Mature particles are presumed to form by budding of the Matrix (M) protein-nucleocapsid complexes through host IMs to acquire host phospholipids and the surface glycoproteins (G). To address mechanisms underlying Nucleorhabdovirus budding, we generated recombinant SYNV G mutants containing a truncated amino-terminal (NT) or carboxyl-terminal (CT) domain. Electron microscopy and sucrose gradient centrifugation analyses showed that the CT domain is essential for virion morphogenesis whereas the NT domain is also required for efficient budding. SYNV infection induces IM invaginations that are thought to provide membrane sites for virus budding. We found that in the context of viral infections, interactions of the M protein with the CT domain of the membrane-anchored G protein mediate M protein translocation and IM invagination. Interestingly, tethering the M protein to endomembranes, either by co-expression with a transmembrane G protein CT domain or by artificial fusion with the G protein membrane targeting sequence, induces IM invagination in uninfected cells. Further evidence to support functions of G-M interactions in virus budding came from dominant negative effects on SYNV-induced IM invagination and viral infections that were elicited by expression of a soluble version of the G protein CT domain. Based on these data, we propose that cooperative G-M interactions promote efficient SYNV budding.
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developments in plant negative strand rna virus reverse genetics
Annual Review of Phytopathology, 2016Co-Authors: Andrew O Jackson, Zhenghe LiAbstract:Twenty years ago, breakthroughs for reverse genetics analyses of negative-strand RNA (NSR) viruses were achieved by devising conditions for generation of infectious viruses in susceptible cells. Recombinant strategies have subsequently been engineered for members of all vertebrate NSR virus families, and research arising from these advances has profoundly increased understanding of infection cycles, pathogenesis, and complexities of host interactions of animal NSR viruses. These strategies also permitted development of many applications, including attenuated vaccines and delivery vehicles for therapeutic and biotechnology proteins. However, for a variety of reasons, it was difficult to devise procedures for reverse genetics analyses of plant NSR viruses. In this review, we discuss advances that have circumvented these problems and resulted in construction of a recombinant system for Sonchus yellow net Nucleorhabdovirus. We also discuss possible extensions to other plant NSR viruses as well as the applicat...
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Developments in Plant Negative-Strand RNA Virus Reverse Genetics.
Annual Review of Phytopathology, 2016Co-Authors: Andrew O JacksonAbstract:Twenty years ago, breakthroughs for reverse genetics analyses of negative-strand RNA (NSR) viruses were achieved by devising conditions for generation of infectious viruses in susceptible cells. Recombinant strategies have subsequently been engineered for members of all vertebrate NSR virus families, and research arising from these advances has profoundly increased understanding of infection cycles, pathogenesis, and complexities of host interactions of animal NSR viruses. These strategies also permitted development of many applications, including attenuated vaccines and delivery vehicles for therapeutic and biotechnology proteins. However, for a variety of reasons, it was difficult to devise procedures for reverse genetics analyses of plant NSR viruses. In this review, we discuss advances that have circumvented these problems and resulted in construction of a recombinant system for Sonchus yellow net Nucleorhabdovirus. We also discuss possible extensions to other plant NSR viruses as well as the applications that may emanate from recombinant analyses of these pathogens.
Zhenghe Li - One of the best experts on this subject based on the ideXlab platform.
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the matrix protein of a plant rhabdovirus mediates superinfection exclusion by inhibiting viral transcription
Journal of Virology, 2019Co-Authors: Xin Zhou, Andrew O Jackson, Zhenghe Li, Xueping ZhouAbstract:Superinfection exclusion (SIE) or cross-protection phenomena have been documented for plant viruses for nearly a century and are widespread among taxonomically diverse viruses, but little information is available about SIE of plant negative-strand RNA viruses. Here, we demonstrate that SIE by sonchus yellow net Nucleorhabdovirus virus (SYNV) is mediated by the viral matrix (M) protein, a multifunctional protein involved in transcription regulation, virion assembly, and virus budding. We show that fluorescent protein-tagged SYNV variants display mutual exclusion/cross-protection in Nicotiana benthamiana plants. Transient expression of the SYNV M protein, but not other viral proteins, interfered with SYNV local infections. In addition, SYNV M deletion mutants failed to exclude superinfection by wild-type SYNV. An SYNV minireplicon reporter gene expression assay showed that the M protein inhibited viral transcription. However, M protein mutants with weakened nuclear localization signals (NLS) and deficient nuclear interactions with the SYNV nucleocapsid protein were unable to suppress transcription. Moreover, SYNV with M NLS mutations exhibited compromised SIE against wild-type SYNV. From these data, we propose that M protein accumulating in nuclei with primary SYNV infections either coils or prevents uncoiling of nucleocapsids released by the superinfecting SYNV virions and suppresses transcription of superinfecting genomes, thereby preventing superinfection. Our model suggests that the rhabdovirus M protein regulates the transition from replication to virion assembly and renders the infected cells nonpermissive for secondary infections.IMPORTANCE Superinfection exclusion (SIE) is a widespread phenomenon in which an established virus infection prevents reinfection by closely related viruses. Understanding the mechanisms governing SIE will not only advance our basic knowledge of virus infection cycles but may also lead to improved design of antiviral measures. Despite the significance of SIE, our knowledge about viral SIE determinants and their modes of actions remain limited. In this study, we show that sonchus yellow net virus (SYNV) SIE is mediated by the viral matrix (M) protein. During primary infections, accumulation of M protein in infected nuclei results in coiling of genomic nucleocapsids and suppression of viral transcription. Consequently, nucleocapsids released by potential superinfectors are sequestered and are unable to initiate new infections. Our data suggest that SYNV SIE is caused by M protein-mediated transition from replication to virion assembly and that this process prevents secondary infections.
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developments in plant negative strand rna virus reverse genetics
Annual Review of Phytopathology, 2016Co-Authors: Andrew O Jackson, Zhenghe LiAbstract:Twenty years ago, breakthroughs for reverse genetics analyses of negative-strand RNA (NSR) viruses were achieved by devising conditions for generation of infectious viruses in susceptible cells. Recombinant strategies have subsequently been engineered for members of all vertebrate NSR virus families, and research arising from these advances has profoundly increased understanding of infection cycles, pathogenesis, and complexities of host interactions of animal NSR viruses. These strategies also permitted development of many applications, including attenuated vaccines and delivery vehicles for therapeutic and biotechnology proteins. However, for a variety of reasons, it was difficult to devise procedures for reverse genetics analyses of plant NSR viruses. In this review, we discuss advances that have circumvented these problems and resulted in construction of a recombinant system for Sonchus yellow net Nucleorhabdovirus. We also discuss possible extensions to other plant NSR viruses as well as the applicat...
Ralf G. Dietzgen - One of the best experts on this subject based on the ideXlab platform.
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Genome-Wide Analysis of Alternative Splicing in Zea mays during Maize Iranian Mosaic Virus Infection
Plant Molecular Biology Reporter, 2019Co-Authors: Abozar Ghorbani, Keramatollah Izadpanah, Alireza Afsharifar, Ahmad Tahmasebi, Ralf G. DietzgenAbstract:Maize Iranian mosaic virus (MIMV) infects several gramineous plants and is an economically important Nucleorhabdovirus in Iran. Maize responds to MIMV infection at the transcriptional level. Alternative splicing (AS) is a mechanism that generates multiple mRNAs from a single pre-mRNA, often encoding protein isoforms with functional differences. We carried out genome-wide analysis of AS responses to MIMV in maize seedlings and identified genes involved in this molecular response. The AS events we investigated included skipped exons, alternative 3′ splice site, alternative 5′ splice site, mutually exclusive exons, and retained introns. In total 10,881 maize genes showed AS, of which 601 genes were involved in response to MIMV-infection and 186 were found only in uninfected maize. AS was identified in some of the genes that are involved in disease resistance or pathogenicity pathways. We demonstrated that in MIMV-infects maize, host genes that are involved in symptom development, virus multiplication, resistance to pathogens and host-pathogen interaction are affected by AS mechanism. Gene network analysis showed that ten genes represent the hubs for the protein network in maize and that they are involved in response to pathogen attack and include 26S proteasome, 14–3-3-like protein A, Rop family, mitogen-activated protein kinase, ubiquitin and serine/threonine-protein kinases. In conclusion, we showed that AS occurs as a transcriptional regulatory mechanism in maize response to MIMV infection and we identified genes that have the key roles in pathogenicity pathways that were differentially spliced in infected seedlings.
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Changes in maize transcriptome in response to maize Iranian mosaic virus infection.
PLOS ONE, 2018Co-Authors: Abozar Ghorbani, Keramatollah Izadpanah, Ralf G. DietzgenAbstract:Background Maize Iranian mosaic virus (MIMV, genus Nucleorhabdovirus, family Rhabdoviridae) causes an economically important disease in maize and other gramineous crops in Iran. MIMV negative-sense RNA genome sequence of 12,426 nucleotides has recently been completed. Maize Genetics and Genomics database shows that 39,498 coding genes and 4,976 non-coding genes of maize have been determined, but still some transcripts could not be annotated. The molecular host cell responses of maize to MIMV infection including differential gene expression have so far not been elucidated. Methodology/principal findings Complementary DNA libraries were prepared from total RNA of MIMV-infected and mock-inoculated maize leaves and sequenced using Illumina HiSeq 2500. Cleaned raw transcript reads from MIMV-infected maize were mapped to reads from uninfected maize and to a maize reference genome. Differentially expressed transcripts were characterized by gene ontology and biochemical pathway analyses. Transcriptome data for selected genes were validated by real-time quantitative PCR. Conclusion/significance Approximately 42 million clean reads for each treatment were obtained. In MIMV-infected maize compared to uninfected plants, 1689 transcripts were up-regulated and 213 transcripts were down-regulated. In response to MIMV infection, several pathways were activated in maize including immune receptor signaling, metabolic pathways, RNA silencing, hormone-mediated pathways, protein degradation, protein kinase and ATP binding activity, and fatty acid metabolism. Also, several transcripts including those encoding hydrophobic protein RCI2B, adenosylmethionine decarboxylase NAC transcription factor and nucleic acid binding, leucine-rich repeat, heat shock protein, 26S proteasome, oxidoreductases and endonuclease activity protein were up-regulated. These data will contribute to the identification of genes and pathways involved in plant-virus interactions that may serve as future targets for improved disease control.
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Gene expression and population polymorphism of maize Iranian mosaic virus in Zea mays, and intracellular localization and interactions of viral N, P, and M proteins in Nicotiana benthamiana
Virus Genes, 2018Co-Authors: Abozar Ghorbani, Keramatollah Izadpanah, Ralf G. DietzgenAbstract:Maize Iranian mosaic virus (MIMV; Mononegavirales , Rhabdoviridae, Nucleorhabdovirus ) infects maize and several other poaceous plants. MIMV encodes six proteins, i.e., nucleocapsid protein (N), polymerase cofactor phosphoprotein (P), putative movement protein (P3), matrix protein (M), glycoprotein (G), and large RNA-dependent RNA polymerase (L). In the present study, MIMV gene expression and genetic polymorphism of an MIMV population in maize were determined. N, P, P3, and M protein genes were more highly expressed than the 5′ terminal G and L genes. Twelve single nucleotide polymorphisms were identified across the genome within a MIMV population in maize from RNA-Seq read data pooled from three infected plants indicating genomic variations of potential importance to evolution of the virus. MIMV N, P, and M proteins that are known to be involved in rhabdovirus replication and transcription were characterized as to their intracellular localization and interactions. N protein accumulated exclusively in the nucleus and interacted with itself and with P protein. P protein accumulated in both the nucleus and cell periphery and interacted with itself, N and M proteins in the nucleus. M protein was localized in the cell periphery and on endomembranes, and interacted with P protein in the nucleus. MIMV proteins show a distinctive combination of intracellular localizations and interactions.
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Completed sequence and corrected annotation of the genome of maize Iranian mosaic virus
Archives of Virology, 2018Co-Authors: Abozar Ghorbani, Keramatollah Izadpanah, Ralf G. DietzgenAbstract:Maize Iranian mosaic virus (MIMV) is a negative-sense single-stranded RNA virus that is classified in the genus Nucleorhabdovirus , family Rhabdoviridae . The MIMV genome contains six open reading frames (ORFs) that encode in 3΄ to 5΄ order the nucleocapsid protein (N), phosphoprotein (P), putative movement protein (P3), matrix protein (M), glycoprotein (G) and RNA-dependent RNA polymerase (L). In this study, we determined the first complete genome sequence of MIMV using Illumina RNA-Seq and 3′/5′ RACE. MIMV genome (‘Fars’ isolate) is 12,426 nucleotides in length. Unexpectedly, the predicted N gene ORF of this isolate and of four other Iranian isolates is 143 nucleotides shorter than that of the MIMV coding-complete reference isolate ‘Shiraz 1’ (Genbank NC_011542), possibly due to a minor error in the previous sequence. Genetic variability among the N, P, P3 and G ORFs of Iranian MIMV isolates was limited, but highest in the G gene ORF. Phylogenetic analysis of complete Nucleorhabdovirus genomes demonstrated a close evolutionary relationship between MIMV, maize mosaic virus and taro vein chlorosis virus.
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Plant rhabdoviruses-their origins and vector interactions.
Current Opinion in Virology, 2018Co-Authors: Anna E. Whitfield, Kathleen M. Martin, Ordom Brian Huot, Hideki Kondo, Ralf G. DietzgenAbstract:Classical plant rhabdoviruses infect monocot and dicot plants, have unsegmented negative-sense RNA genomes and have been taxonomically classified in the genera Cytorhabdovirus and Nucleorhabdovirus. These viruses replicate in their hemipteran vectors and are transmitted in a circulative-propagative mode and virus infection persists for the life of the insect. Based on the discovery of numerous novel rhabdoviruses in arthropods during metagenomic studies and extensive phylogenetic analyses of the family Rhabdoviridae, it is hypothesized that plant-infecting rhabdoviruses are derived from insect viruses. Analyses of viral gene function in plants and insects is beginning to reveal conserved and unique biology for these plant viruses in the two diverse hosts. New tools for insect molecular biology and infectious clones for plant rhabdoviruses are increasing our understanding of the lifestyles of these viruses.
