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Title: 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.
Authors:
 [1] ;  [1] ;  [2] ;  [2] ;  [3]
  1. Instituto de Biologia Molecular y Celular de Plantas (IBMCP), UPV-CSIC, Avda. de los Naranjos, s/n, 46022, Valencia (Spain)
  2. Departament de Bioquimica i Biologia Molecular, Universitat de Valencia, E-46100 Burjassot (Spain)
  3. Instituto de Biologia Molecular y Celular de Plantas (IBMCP), UPV-CSIC, Avda. de los Naranjos, s/n, 46022, Valencia (Spain). E-mail: vpallas@ibmcp.upv.es
Publication Date:
OSTI Identifier:
20729118
Resource Type:
Journal Article
Resource Relation:
Journal Name: Virology; Journal Volume: 339; Journal Issue: 1; Other Information: DOI: 10.1016/j.virol.2005.05.020; PII: S0042-6822(05)00294-1; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; DICHROISM; IN VIVO; MUTANTS; MUTATIONS; PEPTIDES; PLANT CELLS; RNA; VIRUSES