scholarly journals Expression profiling across wild and cultivated tomatoes supports the relevance of early miR482/2118 suppression for Phytophthora resistance

2018 ◽  
Vol 285 (1873) ◽  
pp. 20172560 ◽  
Author(s):  
Sophie de Vries ◽  
Andreas Kukuk ◽  
Janina K. von Dahlen ◽  
Anika Schnake ◽  
Thorsten Kloesges ◽  
...  
Keyword(s):  
Expression Profiling ◽  
Rna Silencing ◽  
Pathogen Resistance ◽  
Tomato Species ◽  
Major Class ◽  
Microrna Family ◽  
Specific Pathogen ◽  
Phytophthora Resistance ◽  
Silencing Suppression ◽  
Leucine Rich Repeats

Plants possess a battery of specific pathogen resistance ( R- )genes. Precise R- gene regulation is important in the presence and absence of a pathogen. Recently, a microRNA family, miR482/2118, was shown to regulate the expression of a major class of R- genes , nucleotide-binding site leucine-rich repeats ( NBS-LRRs ). Furthermore, RNA silencing suppressor proteins, secreted by pathogens, prevent the accumulation of miR482/2118, leading to an upregulation of R- genes. Despite this transcriptional release of R -genes, RNA silencing suppressors positively contribute to the virulence of some pathogens . To investigate this paradox, we analysed how the regulation of NBS-LRRs by miR482/2118 has been shaped by the coevolution between Phytophthora infestans and cultivated and wild tomatoes. We used degradome analyses and qRT-PCR to evaluate and quantify the co-expression of miR482/2118 and their NBS-LRR targets. Our data show that miR482/2118-mediated targeting contributes to the regulation of NBS-LRRs in Solanum lycopersicum. Based on miR482/2118 expression profiling in two additional tomato species—with different coevolutionary histories with P. infestans —we hypothesize that pathogen-mediated RNA silencing suppression is most effective in the interaction between S. lycopersicum and P. infestans . Furthermore, an upregulation of miR482/2118 early in the infection may increase susceptibility to P. infestans .

scholarly journals RNA Silencing Suppression by a Second Copy of the P1 Serine Protease of Cucumber Vein Yellowing Ipomovirus, a Member of the Family Potyviridae That Lacks the Cysteine Protease HCPro

2006 ◽  
Vol 80 (20) ◽  
pp. 10055-10063 ◽  
Author(s):  
Adrian Valli ◽  
Ana Montserrat Martín-Hernández ◽  
Juan José López-Moya ◽  
Juan Antonio García
Keyword(s):  
Rna Silencing ◽  
Fluorescent Protein ◽  
Serine Proteinase ◽  
Plum Pox Virus ◽  
Coding Region ◽  
Long Distance ◽  
Systemic Spread ◽  
The Family ◽  
Silencing Suppression ◽  
Green Fluorescent

ABSTRACT The P1 protein of viruses of the family Potyviridae is a serine proteinase, which is highly variable in length and sequence, and its role in the virus infection cycle is not clear. One of the proposed activities of P1 is to assist HCPro, the product that viruses of the genus Potyvirus use to counteract antiviral defense mediated by RNA silencing. Indeed, an HCPro-coding region is present in all the genomes of members of the genera Potyvirus, Rymovirus, and Tritimovirus that have been sequenced. However, it was recently reported that a sequence coding for HCPro is lacking in the genome of Cucumber vein yellowing virus (CVYV), a member of the genus Ipomovirus, the fourth monopartite genus of the family. In this study, we provide further evidence that P1 enhances the activity of HCPro in members of the genus Potyvirus and show that it is duplicated in the ipomovirus CVYV. The two CVYV P1 copies are arranged in tandem, and the second copy (P1b) has RNA silencing suppression activity. CVYV P1b suppressed RNA silencing induced either by sense green fluorescent protein (GFP) mRNA or by a GFP inverted repeat RNA, indicating that CVYV P1b acts downstream of the formation of double-stranded RNA. CVYV P1b also suppressed local silencing in agroinfiltrated patches of transgenic Nicotiana benthamiana line 16c and delayed its propagation to the neighboring cells. However, neither the short-distance nor long-distance systemic spread of silencing of the GFP transgene was completely blocked by CVYV P1b. CVYV P1b and P1-HCPro from the potyvirus Plum pox virus showed very similar behaviors in all the assays carried out, suggesting that evolution has found a way to counteract RNA silencing by similar mechanisms using very different proteins in viruses of the same family.

scholarly journals Efficient RNA silencing suppression activity of Potato Mop-Top Virus 8K protein is driven by variability and positive selection

Virology ◽  
2019 ◽  
Vol 535 ◽  
pp. 111-121 ◽  
Author(s):  
Pruthvi B. Kalyandurg ◽  
Aminallah Tahmasebi ◽  
Ramesh R. Vetukuri ◽  
Sandeep K. Kushwaha ◽  
Alexander A. Lezzhov ◽  
...  
Keyword(s):  
Positive Selection ◽  
Rna Silencing ◽  
Silencing Suppression ◽  
Potato Mop Top Virus

scholarly journals A Newly Identified Virus in the Family Potyviridae Encodes Two Leader Cysteine Proteases in Tandem That Evolved Contrasting RNA Silencing Suppression Functions

2020 ◽  
Vol 95 (1) ◽  
Author(s):  
Li Qin ◽  
Wentao Shen ◽  
Zhongfa Tang ◽  
Weiyao Hu ◽  
Lingna Shangguan ◽  
...  
Keyword(s):  
Rna Silencing ◽  
Rna Viruses ◽  
Evolutionary Relationship ◽  
Cysteine Proteases ◽  
Terminal Region ◽  
Crop Damage ◽  
Virus Species ◽  
Protease Domain ◽  
The Family ◽  
Silencing Suppression

ABSTRACT Potyviridae is the largest family of plant-infecting RNA viruses and includes many agriculturally and economically important viral pathogens. The viruses in the family, known as potyvirids, possess single-stranded, positive-sense RNA genomes with polyprotein processing as a gene expression strategy. The N-terminal regions of potyvirid polyproteins vary greatly in sequence. Previously, we identified a novel virus species within the family, Areca palm necrotic spindle-spot virus (ANSSV), which was predicted to encode two cysteine proteases, HCPro1 and HCPro2, in tandem at the N-terminal region. Here, we present evidence showing self-cleavage activity of these two proteins and define their cis-cleavage sites. We demonstrate that HCPro2 is a viral suppressor of RNA silencing (VSR), and both the variable N-terminal and conserved C-terminal (protease domain) moieties have antisilencing activity. Intriguingly, the N-terminal region of HCPro1 also has RNA silencing suppression activity, which is, however, suppressed by its C-terminal protease domain, leading to the functional divergence of HCPro1 and HCPro2 in RNA silencing suppression. Moreover, the deletion of HCPro1 or HCPro2 in a newly created infectious clone abolishes viral infection, and the deletion mutants cannot be rescued by addition of corresponding counterparts of a potyvirus. Altogether, these data suggest that the two closely related leader proteases of ANSSV have evolved differential and essential functions to concertedly maintain viral viability. IMPORTANCE The Potyviridae represent the largest group of known plant RNA viruses and account for more than half of the viral crop damage worldwide. The leader proteases of viruses within the family vary greatly in size and arrangement and play key roles during the infection. Here, we experimentally demonstrate the presence of a distinct pattern of leader proteases, HCPro1 and HCPro2 in tandem, in a newly identified member within the family. Moreover, HCPro1 and HCPro2, which are closely related and typically characterized with a short size, have evolved contrasting RNA silencing suppression activity and seem to function in a coordinated manner to maintain viral infectivity. Altogether, the new knowledge fills a missing piece in the evolutionary relationship history of potyvirids and improves our understanding of the diversification of potyvirid genomes.

scholarly journals The Consensus N-Myristoylation Motif of a Geminivirus AC4 Protein Is Required for Membrane Binding and Pathogenicity

2007 ◽  
Vol 20 (4) ◽  
pp. 380-391 ◽  
Author(s):  
Vincent N. Fondong ◽  
R. V. Chowda Reddy ◽  
Cheng Lu ◽  
Bertrand Hankoua ◽  
Christian Felton ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Protein ◽  
Nicotiana Benthamiana ◽  
Rna Silencing ◽  
Membrane Binding ◽  
Confocal Imaging ◽  
East African ◽  
Intrinsic Signals ◽  
Pathogenicity Determinant ◽  
Silencing Suppression

Some geminiviruses encode a small protein, AC4, whose role in pathogenesis has only recently attracted attention. A few studies have shown that this protein is involved in pathogenesis and suppresses RNA silencing. Here, using Nicotiana benthamiana, we show that East African cassava mosaic Cameroon virus (EACMCV) AC4 is a pathogenicity determinant and that it suppresses the systemic phase of RNA silencing. Furthermore, confocal imaging analyses show that it binds preferentially to the plasma membrane as well as to cytosolic membranes including the perinucleus but is excluded from the nucleus. A computational examination of the AC4 protein encoded by the EACMCV, a bipartite geminivirus, shows that it encodes a consensus N-myristoylation motif and is likely posttranslationally myristoylated and palmitoylated. Replacement of Gly-2 and Cys-3 (sites of posttranslational attachment of myristic and palmatic acids, respectively) with alanine affected AC4 membrane binding and pathogenesis. Furthermore, replacement of Ile-5, a nonessential myristoylation residue, with alanine did not affect AC4 function. Together, these data indicate that EACMCV AC4 is likely dually acylated at Gly-2 and Cys-3 and that these modifications are intrinsic signals for membrane targeting and pathogenesis. This is the first report of a membrane protein to be involved in pathogenesis and RNA silencing suppression.

scholarly journals Genetic Analyses of the FRNK Motif Function of Turnip mosaic virus Uncover Multiple and Potentially Interactive Pathways of Cross-Protection

2014 ◽  
Vol 27 (9) ◽  
pp. 944-955 ◽  
Author(s):  
Yi-Jung Kung ◽  
Pin-Chun Lin ◽  
Shyi-Dong Yeh ◽  
Syuan-Fei Hong ◽  
Nam-Hai Chua ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Mosaic Virus ◽  
Rna Silencing ◽  
Turnip Mosaic Virus ◽  
Cross Protection ◽  
Mild Strain ◽  
Silencing Suppression ◽  
Helper Component Proteinase ◽  
Silencing Pathways ◽  
Protection Against Infection

Cross-protection triggered by a mild strain of virus acts as a prophylaxis to prevent subsequent infections by related viruses in plants; however, the underling mechanisms are not fully understood. Through mutagenesis, we isolated a mutant strain of Turnip mosaic virus (TuMV), named Tu-GK, that contains an Arg182Lys substitution in helper component-proteinase (HC-ProK) that confers complete cross-protection against infection by a severe strain of TuMV in Nicotiana benthamiana, Arabidopsis thaliana Col-0, and the Arabidopsis dcl2-4/dcl4-1 double mutant defective in DICER-like ribonuclease (DCL)2/DCL4-mediated silencing. Our analyses showed that HC-ProK loses the ability to interfere with microRNA pathways, although it retains a partial capability for RNA silencing suppression triggered by DCL. We further showed that Tu-GK infection triggers strong salicylic acid (SA)-dependent and SA-independent innate immunity responses. Our data suggest that DCL2/4-dependent and –independent RNA silencing pathways are involved, and may crosstalk with basal innate immunity pathways, in host defense and in cross-protection.

scholarly journals A Single Amino Acid Mutation in the Plum pox virus Helper Component-Proteinase Gene Abolishes Both Synergistic and RNA Silencing Suppression Activities

2005 ◽  
Vol 95 (8) ◽  
pp. 894-901 ◽  
Author(s):  
Pablo González-Jara ◽  
Felix A. Atencio ◽  
Belén Martínez-García ◽  
Daniel Barajas ◽  
Francisco Tenllado ◽  
...  
Keyword(s):  
Amino Acid ◽  
Point Mutation ◽  
Rna Silencing ◽  
Recombinant Virus ◽  
Single Amino Acid ◽  
Single Point Mutation ◽  
Plum Pox Virus ◽  
Helper Component ◽  
Silencing Suppression ◽  
Helper Component Proteinase

The effects on symptom expression of single amino acid mutations in the central region of the Plum pox virus (PPV) helper component-proteinase (HC-Pro) gene were analyzed in Nicotiana benthamiana using Potato virus X (PVX) recombinant viruses. PVX recombinant virus expressing the wild-type variant of PPV HC-Pro induced the expected enhancement of PVX pathogenicity, manifested as necrosis and plant death. Recombinant virus expressing a variant of PPV HC-Pro containing a single point mutation ( HCL134H) was unable to induce this synergistic phenotype. The RNA silencing suppressor activity of PPV HC-Pro was demonstrated in a transient silencing suppression assay. In contrast, the HCL134H mutant showed no such activity. These results indicate that a unique point mutation in PPV HC-Pro impaired its ability to suppress RNA silencing and abolished its capacity to induce synergism, and clearly shows for the first time the link between these two functions in potyvirus HC-Pro. Additionally, we compared the effects on virus accumulation in N. benthamiana plants infected with either the PVX recombinant constructs or with native viruses in double infection experiments. PVX (+) and (-) strand genomic RNA accumulated at similar levels in plants infected with PVX recombinants, leading to an increase in PVX pathology, compared with plants infected with PVX alone. This finding confirms that the enhancement of pathogenicity associated with synergistic interaction is not a consequence of more efficient PVX replication due to RNA silencing suppression by PPV HC-Pro.

scholarly journals The Benyvirus RNA Silencing Suppressor Is Essential for Long-Distance Movement, Requires Both Zinc-Finger and NoLS Basic Residues but Not a Nucleolar Localization for Its Silencing-Suppression Activity

2013 ◽  
Vol 26 (2) ◽  
pp. 168-181 ◽  
Author(s):  
Sotaro Chiba ◽  
Kamal Hleibieh ◽  
Alice Delbianco ◽  
Elodie Klein ◽  
Claudio Ratti ◽  
...  
Keyword(s):  
Zinc Finger ◽  
Rna Silencing ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Nucleolar Localization ◽  
Silencing Suppressor ◽  
Long Distance ◽  
Microscopy Imaging ◽  
Silencing Suppression ◽  
Long Distance Movement

The RNA silencing-suppression properties of Beet necrotic yellow vein virus (BNYVV) and Beet soil-borne mosaic virus (BSBMV) cysteine-rich p14 proteins have been investigated. Suppression of RNA silencing activities were made evident using viral infection of silenced Nicotiana benthamiana 16C, N. benthamiana agroinfiltrated with green fluorescent protein (GFP), and GF-FG hairpin triggers supplemented with viral suppressor of RNA silencing (VSR) constructs or using complementation of a silencing-suppressor-defective BNYVV virus in Chenopodium quinoa. Northern blot analyses of small-interfering RNAs (siRNAs) in agroinfiltration tests revealed reduced amounts of siRNA, especially secondary siRNA, suggesting that benyvirus VSR act downstream of the siRNA production. Using confocal laser-scanning microscopy imaging of infected protoplasts expressing functional p14 protein fused to an enhanced GFP reporter, we showed that benyvirus p14 accumulated in the nucleolus and the cytoplasm independently of other viral factors. Site-directed mutagenesis showed the importance of the nucleolar localization signal embedded in a C4 zinc-finger domain in the VSR function and intrinsic stability of the p14 protein. Conversely, RNA silencing suppression appeared independent of the nucleolar localization of the protein, and a correlation between BNYVV VSR expression and long-distance movement was established.

scholarly journals Translocation of Tomato Bushy Stunt Virus P19 Protein into the Nucleus by ALY Proteins Compromises Its Silencing Suppressor Activity

2006 ◽  
Vol 80 (18) ◽  
pp. 9064-9072 ◽  
Author(s):  
Tomas Canto ◽  
Joachim F. Uhrig ◽  
Maud Swanson ◽  
Kathryn M. Wright ◽  
Stuart A. MacFarlane
Keyword(s):  
Rna Silencing ◽  
Tomato Bushy Stunt Virus ◽  
Bimolecular Fluorescence Complementation ◽  
Rna Recognition Motif ◽  
Rna Recognition ◽  
Long Distance ◽  
Suppressor Activity ◽  
Recognition Motif ◽  
Silencing Suppression ◽  
Suppressor Of Rna Silencing

ABSTRACT The P19 protein of Tomato bushy stunt virus is a potent suppressor of RNA silencing and, depending on the host species, is required for short- and long-distance virus movement and symptom production. P19 interacts with plant ALY proteins and relocalizes a subset of these proteins from the nucleus to the cytoplasm. Here we showed that coexpression by agroinfiltration in Nicotiana benthamiana of P19 and the subset of ALY proteins that are not relocalized from the nucleus interfered with the ability of P19 to suppress RNA silencing. We demonstrated that this interference correlates with the relocation of P19 from the cytoplasm into the nucleus, and by constructing and analyzing chimeric ALY genes, we showed that the C-terminal part of the central, RNA recognition motif of ALY is responsible for interaction with P19, relocalization or nonrelocalization of ALY, and inhibition of silencing suppression by P19. We studied the interaction of ALY and P19 by using the technique of bimolecular fluorescence complementation to show that these proteins associate physically in the nucleus but not detectably in the cytoplasm, and we present a model to explain the dynamics of this interaction.

scholarly journals Symptom induction and RNA silencing suppression by the cucumber mosaic virus 2b protein

2010 ◽  
Vol 5 (6) ◽  
pp. 705-708 ◽  
Author(s):  
Mathew G. Lewsey ◽  
Inmaculada González ◽  
Natalia O. Kalinina ◽  
Peter Palukaitis ◽  
Tomas Canto ◽  
...  
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Rna Silencing ◽  
2B Protein ◽  
Silencing Suppression

scholarly journals The Cucumber vein yellowing virus Silencing Suppressor P1b Can Functionally Replace HCPro in Plum pox virus Infection in a Host-Specific Manner

2012 ◽  
Vol 25 (2) ◽  
pp. 151-164 ◽  
Author(s):  
Alberto Carbonell ◽  
Gabriela Dujovny ◽  
Juan Antonio García ◽  
Adrian Valli
Keyword(s):  
Rna Silencing ◽  
Deleterious Effect ◽  
Prunus Persica ◽  
Plant Viruses ◽  
Natural Host ◽  
Plum Pox Virus ◽  
Silencing Suppressor ◽  
Specific Manner ◽  
Silencing Suppression ◽  
Yellowing Virus

Plant viruses of the genera Potyvirus and Ipomovirus (Potyviridae family) use unrelated RNA silencing suppressors (RSS) to counteract antiviral RNA silencing responses. HCPro is the RSS of Potyvirus spp., and its activity is enhanced by the upstream P1 protein. Distinctively, the ipomovirus Cucumber vein yellowing virus (CVYV) lacks HCPro but contains two P1 copies in tandem (P1aP1b), the second of which functions as RSS. Using chimeras based on the potyvirus Plum pox virus (PPV), we found that P1b can functionally replace HCPro in potyviral infections of Nicotiana plants. Interestingly, P1a, the CVYV protein homologous to potyviral P1, disrupted the silencing suppression activity of P1b and reduced the infection efficiency of PPV in Nicotiana benthamiana. Testing the influence of RSS in host specificity, we found that a P1b-expressing chimera poorly infected PPV's natural host, Prunus persica. Conversely, P1b conferred on PPV chimeras the ability to replicate locally in cucumber, CVYV's natural host. The deleterious effect of P1a on PPV infection is host dependent, because the P1aP1b-expressing PPV chimera accumulated in cucumber to higher levels than PPV expressing P1b alone. These results demonstrate that a potyvirus can use different RSS, and that particular RSS and upstream P1-like proteins contribute to defining the virus host range.

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