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1 Département de Virologie, Institut de Biologie Moléculaire des Plantes du CNRS, 12 rue du Général Zimmer, 67084 Strasbourg, France
2 Fakultät für Biologie, Lehrstuhl für Genomforschung, 33594 Bielefeld, Germany
Correspondence
David Gilmer
david.gilmer{at}ibmp-ulp.u-strasbg.fr
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-
. 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.
Present address: LBMAGM, 42 rue du laboratoire, L1011 Luxembourg.
Present address: IBVM INRA, 71 Avenue E. Bourleaux, BP81, 33883 Villenave d'Ornon Cedex, France.
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