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Renato O Resende - One of the best experts on this subject based on the ideXlab platform.
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The NSm proteins of phylogenetically related tospoviruses trigger Sw-5b–mediated resistance dissociated of their Cell-to-Cell Movement function
Virus Research, 2017Co-Authors: Mikhail O. Leastro, Vicente Pallás, Athos Silva De Oliveira, Jesús A. Sánchez-navarro, Richard Kormelink, Renato O ResendeAbstract:Abstract The Cell-to-Cell Movement protein (NSM) of tomato spotted wilt virus (TSWV) has been recently identified as the effector of the single dominant Sw-5b resistance gene from tomato (Solanum lycopersicum L.). Although most TSWV isolates shows a resistance-inducing (RI) phenotype, regular reports have appeared on the emergence of resistance-breaking (RB) isolates in tomato fields, and suggested a strong association with two point mutations (C118Y and T120N) in the NSM protein. In this study the Sw-5b gene has been demonstrated to confer not only resistance against TSWV but to members of five additional, phylogenetically-related classified within the so-called “American” evolutionary clade, i.e., Alstroemeria necrotic streak virus (ANSV), chrysanthemum stem necrosis virus (CSNV), groundnut ringspot virus (GRSV), Impatiens necrotic spot virus (INSV) and tomato chlorotic spot virus (TCSV). Remarkably, bean necrotic mosaic virus (BeNMV), a recently discovered tospovirus classified in a distinct American subclade and circulating on the American continent, did not trigger a Sw-5b-mediated hypersensitive (HR) response. Introduction of point mutations C118Y and T120N into the NSM protein of TSWV, TCSV and CSNV abrogated the ability to trigger Sw-5b-mediated HR in both transgenic-N. benthamiana and tomato isolines harboring the Sw-5b gene whereas it had no effect on BeNMV NSM. Truncated versions of TSWV NSM lacking motifs associated with tubule formation, Cell-to-Cell or systemic viral Movement were made and tested for triggering of resistance. HR was still observed with truncated NSM proteins lacking 50 amino acids (out of 301) from either the amino- or carboxy-terminal end. These data altogether indicate the importance of amino acid residues C118 and T120 in Sw-5b-mediated HR only for the NSM proteins from one cluster of tospoviruses within the American clade, and that the ability to support viral Cell-to-Cell Movement is not required for effector functionality.
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the tomato spotted wilt virus Cell to Cell Movement protein nsm triggers a hypersensitive response in sw 5 containing resistant tomato lines and in nicotiana benthamiana transformed with the functional sw 5b resistance gene copy
Molecular Plant Pathology, 2014Co-Authors: Mariana Hallwass, Athos Silva De Oliveira, Erico De Campos Dianese, Dick Lohuis, L S Boiteux, Alice K Inouenagata, Renato O ResendeAbstract:Although the Sw-5 gene cluster has been cloned, and Sw-5b has been identified as the functional gene copy that confers resistance to Tomato spotted wilt virus (TSWV), its avirulence (Avr) determinant has not been identified to date. Nicotiana tabacum 'SR1' plants transformed with a copy of the Sw-5b gene are immune without producing a clear visual response on challenge with TSWV, whereas it is shown here that N. benthamiana transformed with Sw-5b gives a rapid and conspicuous hypersensitive response (HR). Using these plants, from all structural and non-structural TSWV proteins tested, the TSWV Cell-to-Cell Movement protein (NSM ) was confirmed as the Avr determinant using a Potato virus X (PVX) replicon or a non-replicative pEAQ-HT expression vector system. HR was induced in Sw-5b-transgenic N. benthamiana as well as in resistant near-isogenic tomato lines after agroinfiltration with a functional Cell-to-Cell Movement protein (NSM ) from a resistance-inducing (RI) TSWV strain (BR-01), but not with NSM from a Sw-5 resistance-breaking (RB) strain (GRAU). This is the first biological demonstration that Sw-5-mediated resistance is triggered by the TSWV NSM Cell-to-Cell Movement protein.
Athos Silva De Oliveira - One of the best experts on this subject based on the ideXlab platform.
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The NSm proteins of phylogenetically related tospoviruses trigger Sw-5b–mediated resistance dissociated of their Cell-to-Cell Movement function
Virus Research, 2017Co-Authors: Mikhail O. Leastro, Vicente Pallás, Athos Silva De Oliveira, Jesús A. Sánchez-navarro, Richard Kormelink, Renato O ResendeAbstract:Abstract The Cell-to-Cell Movement protein (NSM) of tomato spotted wilt virus (TSWV) has been recently identified as the effector of the single dominant Sw-5b resistance gene from tomato (Solanum lycopersicum L.). Although most TSWV isolates shows a resistance-inducing (RI) phenotype, regular reports have appeared on the emergence of resistance-breaking (RB) isolates in tomato fields, and suggested a strong association with two point mutations (C118Y and T120N) in the NSM protein. In this study the Sw-5b gene has been demonstrated to confer not only resistance against TSWV but to members of five additional, phylogenetically-related classified within the so-called “American” evolutionary clade, i.e., Alstroemeria necrotic streak virus (ANSV), chrysanthemum stem necrosis virus (CSNV), groundnut ringspot virus (GRSV), Impatiens necrotic spot virus (INSV) and tomato chlorotic spot virus (TCSV). Remarkably, bean necrotic mosaic virus (BeNMV), a recently discovered tospovirus classified in a distinct American subclade and circulating on the American continent, did not trigger a Sw-5b-mediated hypersensitive (HR) response. Introduction of point mutations C118Y and T120N into the NSM protein of TSWV, TCSV and CSNV abrogated the ability to trigger Sw-5b-mediated HR in both transgenic-N. benthamiana and tomato isolines harboring the Sw-5b gene whereas it had no effect on BeNMV NSM. Truncated versions of TSWV NSM lacking motifs associated with tubule formation, Cell-to-Cell or systemic viral Movement were made and tested for triggering of resistance. HR was still observed with truncated NSM proteins lacking 50 amino acids (out of 301) from either the amino- or carboxy-terminal end. These data altogether indicate the importance of amino acid residues C118 and T120 in Sw-5b-mediated HR only for the NSM proteins from one cluster of tospoviruses within the American clade, and that the ability to support viral Cell-to-Cell Movement is not required for effector functionality.
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the tomato spotted wilt virus Cell to Cell Movement protein nsm triggers a hypersensitive response in sw 5 containing resistant tomato lines and in nicotiana benthamiana transformed with the functional sw 5b resistance gene copy
Molecular Plant Pathology, 2014Co-Authors: Mariana Hallwass, Athos Silva De Oliveira, Erico De Campos Dianese, Dick Lohuis, L S Boiteux, Alice K Inouenagata, Renato O ResendeAbstract:Although the Sw-5 gene cluster has been cloned, and Sw-5b has been identified as the functional gene copy that confers resistance to Tomato spotted wilt virus (TSWV), its avirulence (Avr) determinant has not been identified to date. Nicotiana tabacum 'SR1' plants transformed with a copy of the Sw-5b gene are immune without producing a clear visual response on challenge with TSWV, whereas it is shown here that N. benthamiana transformed with Sw-5b gives a rapid and conspicuous hypersensitive response (HR). Using these plants, from all structural and non-structural TSWV proteins tested, the TSWV Cell-to-Cell Movement protein (NSM ) was confirmed as the Avr determinant using a Potato virus X (PVX) replicon or a non-replicative pEAQ-HT expression vector system. HR was induced in Sw-5b-transgenic N. benthamiana as well as in resistant near-isogenic tomato lines after agroinfiltration with a functional Cell-to-Cell Movement protein (NSM ) from a resistance-inducing (RI) TSWV strain (BR-01), but not with NSM from a Sw-5 resistance-breaking (RB) strain (GRAU). This is the first biological demonstration that Sw-5-mediated resistance is triggered by the TSWV NSM Cell-to-Cell Movement protein.
Vicente Pallás - One of the best experts on this subject based on the ideXlab platform.
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A conserved motif in three viral Movement proteins from different genera is required for host factor recruitment and Cell-to-Cell Movement
Scientific reports, 2020Co-Authors: José A. Navarro, Marta Serra-soriano, Lorena Corachán-valencia, Vicente PallásAbstract:Due to their minimal genomes, plant viruses are forced to hijack specific Cellular pathways to ensure host colonization, a condition that most frequently involves physical interaction between viral and host proteins. Among putative viral interactors are the Movement proteins, responsible for plasmodesma gating and genome binding during viral transport. Two of them, DGBp1 and DGBp2, are required for alpha-, beta- and gammacarmovirus Cell-to-Cell Movement, but the number of DGBp-host interactors identified at present is limited. By using two different approaches, yeast two-hybrid and bimolecular fluorescence complementation assays, we found three Arabidopsis factors, eIF3g1, RPP3A and WRKY36, interacting with DGBp1s from each genus mentioned above. eIF3g1 and RPP3A are mainly involved in protein translation initiation and elongation phases, respectively, while WRKY36 belongs to WRKY transcription factor family, important regulators of many defence responses. These host proteins are not expected to be associated with viral Movement, but knocking out WRKY36 or silencing either RPP3A or eIF3g1 negatively affected Arabidopsis infection by Turnip crinkle virus. A highly conserved FNF motif at DGBp1 C-terminus was required for protein-protein interaction and Cell-to-Cell Movement, suggesting an important biological role.
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The NSm proteins of phylogenetically related tospoviruses trigger Sw-5b–mediated resistance dissociated of their Cell-to-Cell Movement function
Virus Research, 2017Co-Authors: Mikhail O. Leastro, Vicente Pallás, Athos Silva De Oliveira, Jesús A. Sánchez-navarro, Richard Kormelink, Renato O ResendeAbstract:Abstract The Cell-to-Cell Movement protein (NSM) of tomato spotted wilt virus (TSWV) has been recently identified as the effector of the single dominant Sw-5b resistance gene from tomato (Solanum lycopersicum L.). Although most TSWV isolates shows a resistance-inducing (RI) phenotype, regular reports have appeared on the emergence of resistance-breaking (RB) isolates in tomato fields, and suggested a strong association with two point mutations (C118Y and T120N) in the NSM protein. In this study the Sw-5b gene has been demonstrated to confer not only resistance against TSWV but to members of five additional, phylogenetically-related classified within the so-called “American” evolutionary clade, i.e., Alstroemeria necrotic streak virus (ANSV), chrysanthemum stem necrosis virus (CSNV), groundnut ringspot virus (GRSV), Impatiens necrotic spot virus (INSV) and tomato chlorotic spot virus (TCSV). Remarkably, bean necrotic mosaic virus (BeNMV), a recently discovered tospovirus classified in a distinct American subclade and circulating on the American continent, did not trigger a Sw-5b-mediated hypersensitive (HR) response. Introduction of point mutations C118Y and T120N into the NSM protein of TSWV, TCSV and CSNV abrogated the ability to trigger Sw-5b-mediated HR in both transgenic-N. benthamiana and tomato isolines harboring the Sw-5b gene whereas it had no effect on BeNMV NSM. Truncated versions of TSWV NSM lacking motifs associated with tubule formation, Cell-to-Cell or systemic viral Movement were made and tested for triggering of resistance. HR was still observed with truncated NSM proteins lacking 50 amino acids (out of 301) from either the amino- or carboxy-terminal end. These data altogether indicate the importance of amino acid residues C118 and T120 in Sw-5b-mediated HR only for the NSM proteins from one cluster of tospoviruses within the American clade, and that the ability to support viral Cell-to-Cell Movement is not required for effector functionality.
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contribution of topology determinants of a viral Movement protein to its membrane association intraCellular traffic and viral Cell to Cell Movement
Journal of Virology, 2011Co-Authors: Ainhoa Genoves, Vicente Pallás, Joseantonio NavarroAbstract:The p7B Movement protein (MP) of Melon necrotic spot virus (MNSV) is a single-pass membrane protein associated with the endoplasmic reticulum (ER), the Golgi apparatus (GA), and plasmodesmata (Pd). Experimental data presented here revealed that the p7B transmembrane domain (TMD) was sufficient to target the green fluorescent protein (GFP) to ER membranes. In addition, the short extramembrane regions of p7B were essential for subsequent ER export and transport to the GA and Pd. Microsomal partitioning and bimolecular fluorescence assays supported a type II topology of p7B in planta. Mutations affecting conventional determinants of p7B membrane topology, such as the TMD secondary structure, the overall hydrophobicity profile, the so-called “aromatic belt,” and the net charge distribution on either side of the TMD, were engineered into infectious RNAs to investigate the relationship between the MP structure and MNSV Cell-to-Cell Movement. The results revealed that (i) the overall hydrophobic profile and the α-helix integrity of the TMD were relevant for virus Movement, (ii) modification of the net charge balance of the regions flanking both TMD sides drastically reduced Cell-to-Cell Movement, (iii) localization of p7B to the GA was necessary but not sufficient for virus Movement, and (iv) membrane insertion was essential for p7B function in virus Movement. Our results therefore indicate that MNSV Cell-to-Cell Movement requires sequential transport of p7B from the ER via the GA to Pd, which is modulated by a combination of several signals with different strengths in the extramembrane regions and TMD of the MP.
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a self interacting carmovirus Movement protein plays a role in binding of viral rna during the Cell to Cell Movement and shows an actin cytoskeleton dependent location in Cell periphery
Virology, 2009Co-Authors: Ainhoa Genoves, Joseantonio Navarro, Vicente PallásAbstract:The p7A of Melon necrotic spot virus has been described to be a RNA-binding Movement protein essential for Cell-to-Cell Movement but its role in this process is still unknown. Here, we found that primary and secondary structure elements on p7A appear to form a composite RNA-binding site required for both RNA interaction and Cell-to-Cell Movement in plants indicating a direct correlation between these activities. Furthermore, we found that fluorescent-tagged p7A was distributed in punctuate structures at the Cell periphery but also in motile cytoplasmic inclusion bodies which were in close association with the actin MFs and most likely generated by self-interacting p7A molecules as shown by BiFC assays. Consistently, the p7A subCellular distribution was revealed to be sensitive to the actin inhibitor, latrunculin B. The involvement of the RNA-binding capabilities and the subCellular location of the p7A in the intraCellular and interCellular virus Movement is discussed.
Susan M Angell - One of the best experts on this subject based on the ideXlab platform.
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tip a novel host factor linking callose degradation with the Cell to Cell Movement of potato virus x
Molecular Plant-microbe Interactions, 2003Co-Authors: Ingela Fridborg, Jef Grainger, Anthony Page, Mark Coleman, Kim Findlay, Susan M AngellAbstract:The Cell-to-Cell Movement of Potato virus X (PVX) requires four virus-encoded proteins, the triple gene block (TGB) proteins (TGB25K, TGB12K, and TGB8K) and the coat protein. TGB12K increases the plasmodesmal size exclusion limit (SEL) and may, therefore, interact directly with components of the Cell wall or with plant proteins associated with bringing about this change. A yeast two-hybrid screen using TGB12K as bait identified three TGB12K-interacting proteins (TIP1, TIP2, and TIP3). All three TIPs interacted specifically with TGB12K but not with TGB25K or TGB8K. Similarly, all three TIPs interacted with β-1,3-glucanase, the enzyme that may regulate plasmodesmal SEL through callose degradation. Sequence analyses revealed that the TIPs encode very similar proteins and that TIP1 corresponds to the tobacco ankyrin repeat-containing protein HBP1. A TIP1::GFP fusion protein localized to the cytoplasm. Coexpression of this fusion protein with TGB12K induced Cellular changes manifested as deposits of additional...
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tip a novel host factor linking callose degradation with the Cell to Cell Movement of potato virus x
Molecular Plant-microbe Interactions, 2003Co-Authors: Ingela Fridborg, Jef Grainger, Anthony Page, Mark Coleman, Kim Findlay, Susan M AngellAbstract:The Cell-to-Cell Movement of Potato virus X (PVX) requires four virus-encoded proteins, the triple gene block (TGB) proteins (TGB25K, TGB12K, and TGB8K) and the coat protein. TGB12K increases the plasmodesmal size exclusion limit (SEL) and may, therefore, interact directly with components of the Cell wall or with plant proteins associated with bringing about this change. A yeast two-hybrid screen using TGB12K as bait identified three TGB12K-interacting proteins (TIP1, TIP2, and TIP3). All three TIPs interacted specifically with TGB12K but not with TGB25K or TGB8K. Similarly, all three TIPs interacted with beta-1,3-glucanase, the enzyme that may regulate plasmodesmal SEL through callose degradation. Sequence analyses revealed that the TIPs encode very similar proteins and that TIP1 corresponds to the tobacco ankyrin repeat-containing protein HBP1. A TIP1::GFP fusion protein localized to the cytoplasm. Coexpression of this fusion protein with TGB12K induced Cellular changes manifested as deposits of additional cytoplasm at the Cell periphery. This work reports a direct link between a viral Movement protein required to increase plasmodesmal SEL and a host factor that has been implicated as a key regulator of plasmodesmal SEL. We propose that the TIPs are susceptibility factors that modulate the plasmodesmal SEL.
Mariana Hallwass - One of the best experts on this subject based on the ideXlab platform.
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the tomato spotted wilt virus Cell to Cell Movement protein nsm triggers a hypersensitive response in sw 5 containing resistant tomato lines and in nicotiana benthamiana transformed with the functional sw 5b resistance gene copy
Molecular Plant Pathology, 2014Co-Authors: Mariana Hallwass, Athos Silva De Oliveira, Erico De Campos Dianese, Dick Lohuis, L S Boiteux, Alice K Inouenagata, Renato O ResendeAbstract:Although the Sw-5 gene cluster has been cloned, and Sw-5b has been identified as the functional gene copy that confers resistance to Tomato spotted wilt virus (TSWV), its avirulence (Avr) determinant has not been identified to date. Nicotiana tabacum 'SR1' plants transformed with a copy of the Sw-5b gene are immune without producing a clear visual response on challenge with TSWV, whereas it is shown here that N. benthamiana transformed with Sw-5b gives a rapid and conspicuous hypersensitive response (HR). Using these plants, from all structural and non-structural TSWV proteins tested, the TSWV Cell-to-Cell Movement protein (NSM ) was confirmed as the Avr determinant using a Potato virus X (PVX) replicon or a non-replicative pEAQ-HT expression vector system. HR was induced in Sw-5b-transgenic N. benthamiana as well as in resistant near-isogenic tomato lines after agroinfiltration with a functional Cell-to-Cell Movement protein (NSM ) from a resistance-inducing (RI) TSWV strain (BR-01), but not with NSM from a Sw-5 resistance-breaking (RB) strain (GRAU). This is the first biological demonstration that Sw-5-mediated resistance is triggered by the TSWV NSM Cell-to-Cell Movement protein.