Expression dynamics and ultrastructural localization of epitope-tagged Abutilon mosaic virus nuclear shuttle and movement proteins inNicotiana benthamianacells

The geminivirus Abutilon mosaic virus (AbMV)encodes two proteins which are essential for viral spread within plants. The nuclear shuttle protein (NSP)transfers viral DNA between the nucleus and cytoplasm, whereas the movement protein (MP)facilitates transport between cells through plasmodesmata and long-distance via phloem. An inducible overexpression system for epitope-tagged NSP and MP in plants yielded unprecedented amounts of both proteins. Western blots revealed extensive posttranslational modification and truncation for MP, but not for NSP. Ultrastructural examination ofNicotiana benthamianatissues showed characteristic nucleopathic alterations, including fibrillar rings, when epitope-tagged NSP and MP were simultaneously expressed in leaves locally infected with an AbMV DNA A in which the coat protein gene was replaced by a green fluorescent protein encoding gene. Immunogold labelling localized NSP in the nucleoplasm and in thefibrillar rings. MP appeared at the cell periphery, probably the plasma membrane, and plasmodesmata.