Nucleo-cytoplasmic shuttling of the beet necrotic yellow vein virus RNA-3-encoded p25 protein.
The protein p25 encoded by beet necrotic yellow vein virus (BNYVV) RNA-3 is involved in symptom expression of infected plants. Confocal microscopy analysis of wild-type and mutated p25 fused to GFP and transiently expressed in BY-2 tobacco suspension cells identified a nuclear localization signal (NLS) in the N-terminal part of the protein. Functionality of the NLS was confirmed by pull-down assays using rice and pepper importin-alpha. Furthermore, it was demonstrated that p25 contains a nuclear export sequence sensitive to leptomycin B. The nuclear export signal (NES) was characterized by mutagenesis. A GFP-p25 fusion protein expressed during a BNYVV infection of Chenopodium quinoa leaves had the same subcellular localization as observed during transient expression in BY-2 cells. The symptom phenotype induced by expression of GFP-p25 during infection was similar to that induced by wild-type virus. Studies with mutated derivatives of GFP-p25 revealed that symptom phenotype was altered when the subcellular localization of GFP-p25 was modified.
Vetter
Guillaume
G
Département de Virologie, Institut de Biologie Moléculaire des Plantes du CNRS, 12 rue du Général Zimmer, 67084 Strasbourg, France.
Hily
Jean-Michel
JM
Département de Virologie, Institut de Biologie Moléculaire des Plantes du CNRS, 12 rue du Général Zimmer, 67084 Strasbourg, France.
Klein
Elodie
E
Département de Virologie, Institut de Biologie Moléculaire des Plantes du CNRS, 12 rue du Général Zimmer, 67084 Strasbourg, France.
Schmidlin
Laure
L
Département de Virologie, Institut de Biologie Moléculaire des Plantes du CNRS, 12 rue du Général Zimmer, 67084 Strasbourg, France.
Haas
Muriel
M
Département de Virologie, Institut de Biologie Moléculaire des Plantes du CNRS, 12 rue du Général Zimmer, 67084 Strasbourg, France.
Merkle
Thomas
T
Fakultät für Biologie, Lehrstuhl für Genomforschung, 33594 Bielefeld, Germany.
Gilmer
David
D
Département de Virologie, Institut de Biologie Moléculaire des Plantes du CNRS, 12 rue du Général Zimmer, 67084 Strasbourg, France.
D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D027461:Beta vulgaris / Q000821:virology*; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D028884:Karyopherins / Q000378:metabolism; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D010942:Plant Viruses / Q000737:chemistry*; D012328:RNA Viruses / Q000737:chemistry*; D014162:Transfection; D014764:Viral Proteins / Q000378:metabolism*
D028884:Karyopherins; D019913:Nuclear Localization Signals; D014764:Viral Proteins