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.
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.
Laboratory of Plant Molecular Virology, Instituto de Biología Molecular y Celular de Plantas, IBMCP (Universitat Politècnica de València-Consejo Superior de Investigaciones Científicas), Ingeniero Fausto Elio s/n, Valencia, 46022, Spain.
Marta
M
Serra-Soriano
Laboratory of Plant Molecular Virology, Instituto de Biología Molecular y Celular de Plantas, IBMCP (Universitat Politècnica de València-Consejo Superior de Investigaciones Científicas), Ingeniero Fausto Elio s/n, Valencia, 46022, Spain.
José
J
Antonio Navarro
Laboratory of Plant Molecular Virology, Instituto de Biología Molecular y Celular de Plantas, IBMCP (Universitat Politècnica de València-Consejo Superior de Investigaciones Científicas), Ingeniero Fausto Elio s/n, Valencia, 46022, Spain.
Vicente
V
Pallás
D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D020868:Gene Silencing; D010942:Plant Viruses / Q000235:genetics / Q000472:pathogenicity; D017863:Potexvirus / Q000235:genetics / Q000472:pathogenicity; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*
D036022:Capsid Proteins; D012367:RNA, Viral; C090159:coat protein, Melon necrotic spot virus
RNA binding; coat protein; plant virus; silencing