Subcellular sorting of small membrane-associated triple gene block proteins: TGBp3-assisted targeting of TGBp2

We studied subcellular distribution of green fluorescent protein (GFP)-tagg ed movement proteins encoded by the second and the third genes of poa semil atent hordeivirus (PSLV) triple gene block (TGB), 15K TGBp2 and 18K TGBp3. GFP-15K transiently expressed in Nicotiana benthamiana leaf epidermal cells was associated with the endomembrane system elements. GFP-18K appeared in the membrane bodies at cell periphery. Mutation analysis demonstrated that subcellular targeting of GFP-15K depended on the protein transmembrane segm ent(s), whereas the TGBp3 central hydrophilic region was responsible for ta rgeting of GFP-18K. Coexpression of GFP-15K with the intact 18K protein ind uced drastic changes in the TGBp2 localization: GFP-15K appeared in the cel l peripheral bodies similar to those in the cells expressing GFP-18K alone. Coexpression experiments with mutant forms of both proteins argue against involvement of direct interaction between small TGB proteins in the TGBp3-a ssisted targeting of TGBp2 to the cell peripheral compartments. This conclu sion was further confirmed by similar effects on the PSLV 15K TGBp2 localiz ation induced by TGBp3 proteins of PSLV and potato virus X, which have no d etectable sequence similarity to each other. (C) 2000 Academic Press.