Mutational analysis of the RNA-binding domain of the Prunus necrotic ringspot virus (PNRSV) movement protein reveals its requirement for cell-to-cell movement.
The movement protein (MP) of Prunus necrotic ringspot virus (PNRSV) is required for cell-to-cell movement. MP subcellular localization studies using a GFP fusion protein revealed highly punctate structures between neighboring cells, believed to represent plasmodesmata. Deletion of the RNA-binding domain (RBD) of PNRSV MP abolishes the cell-to-cell movement. A mutational analysis on this RBD was performed in order to identify in vivo the features that govern viral transport. Loss of positive charges prevented the cell-to-cell movement even though all mutants showed a similar accumulation level in protoplasts to those observed with the wild-type (wt) MP. Synthetic peptides representing the mutants and wild-type RBDs were used to study RNA-binding affinities by EMSA assays being approximately 20-fold lower in the mutants. Circular dichroism analyses revealed that the secondary structure of the peptides was not significantly affected by mutations. The involvement of the affinity changes between the viral RNA and the MP in the viral cell-to-cell movement is discussed.
Carmen Herranz
Ma
M
Instituto de Biología Molecular y Celular de Plantas (IBMCP), UPV-CSIC, Avda. de los Naranjos, s/n, 46022, Valencia, Spain.
Sanchez-Navarro
Jesús-Angel
JA
Saurí
Ana
A
Mingarro
Ismael
I
Pallás
Vicente
V
D000595:Amino Acid Sequence; D001692:Biological Transport; D019179:Ilarvirus / Q000378:metabolism*; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000821:virology; D053830:Plant Viral Movement Proteins; D017434:Protein Structure, Tertiary / Q000235:genetics; D011523:Protoplasts / Q000821:virology; D012367:RNA, Viral / Q000378:metabolism*; D014026:Nicotiana / Q000821:virology; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
D053830:Plant Viral Movement Proteins; D012367:RNA, Viral; D014764:Viral Proteins