A Protocol to Measure the Extent of Cell-to-cell Movement of RNA Viruses in Planta

Navarro, J. A.; Serra-Soriano, Marta; Pallás, Vicente

doi:10.21769/bioprotoc.1269

Cited by 2 publications

(3 citation statements)

References 2 publications

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“…Cell‐to‐cell movement was examined as described previously by monitoring the formation of fluorescent foci by MNSV(Al/GFP) constructs (Navarro et al ., ; Serra‐Soriano et al ., ). Binary constructs were agroinfiltrated [optical density at 600 nm (OD 600 ) of 0.2] into fully expanded leaves from 2‐week‐old melon plants.…”

Section: Methodsmentioning

confidence: 99%

Dissecting the multifunctional role of the N‐terminal domain of the Melon necrotic spot virus coat protein in RNA packaging, viral movement and interference with antiviral plant defence

Serra-Soriano

Navarro

Pallás

2016

Molecular Plant Pathology

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The coat protein (CP) of Melon necrotic spot virus (MNSV) is structurally composed of three major domains. The middle S-domain builds a robust protein shell around the viral genome, whereas the C-terminal protruding domain, or P-domain, is involved in the attachment of virions to the transmission vector. Here, we have shown that the N-terminal domain, or R-domain, and the arm region, which connects the R-domain and S-domain, are involved in different key steps of the viral cycle, such as cell-to-cell movement and the suppression of RNA silencing and pathogenesis through their RNA-binding capabilities. Deletion mutants revealed that the CP RNA-binding ability was abolished only after complete, but not partial, deletion of the R-domain and the arm region. However, a comparison of the apparent dissociation constants for the CP RNA-binding reaction of several partial deletion mutants showed that the arm region played a more relevant role than the R-domain in in vitro RNA binding. Similar results were obtained in in vivo assays, although, in this case, full-length CPs were required to encapsidate full-length genomes. We also found that the R-domain carboxyl portion and the arm region were essential for efficient cell-to-cell movement, for enhancement of Potato virus X pathogenicity, for suppression of systemic RNA silencing and for binding of small RNAs. Therefore, unlike other carmovirus CPs, the R-domain and the arm region of MNSV CP have acquired, in addition to other essential functions such as genome binding and encapsidation functions, the ability to suppress RNA silencing by preventing systemic small RNA transport.

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Section: Methodsmentioning

confidence: 99%

Dissecting the multifunctional role of the N‐terminal domain of the Melon necrotic spot virus coat protein in RNA packaging, viral movement and interference with antiviral plant defence

Serra-Soriano

Navarro

Pallás

2016

Molecular Plant Pathology

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“…GraphPad Prism 6 software and Student's t-test (p value<0.05) was used to determine the statistical significance of results. For a detailed description of the protocol see also (Navarro et al, 2014).…”

Section: Plasmid Construction For Cell-to-cell Movement Assays and Ex...mentioning

confidence: 99%

“…Cell-to-cell movement was monitored at 6 and 12 days post-infiltration (dpi) by measuring the area of fluorescent infection foci in six fully expanded melon leaves per mutant (Navarro et al, 2014;Serra-Soriano et al, 2014). Each bar diagram in Fig.…”

Section: The R Domain Of Mnsv Cp Modulates Cell-to-cell Movement Inde...mentioning

confidence: 99%

Relación estructura-función de las proteínas virales implicadas en el movimiento de los carmovirus y su interacción con factores celulares

Soriano¹

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A Protocol to Measure the Extent of Cell-to-cell Movement of RNA Viruses in Planta

Cited by 2 publications

References 2 publications

Dissecting the multifunctional role of the N‐terminal domain of the Melon necrotic spot virus coat protein in RNA packaging, viral movement and interference with antiviral plant defence

Dissecting the multifunctional role of the N‐terminal domain of the Melon necrotic spot virus coat protein in RNA packaging, viral movement and interference with antiviral plant defence

Relación estructura-función de las proteínas virales implicadas en el movimiento de los carmovirus y su interacción con factores celulares

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