Subcellular Sorting of Small Membrane-Associated Triple Gene Block Proteins: TGBp3-Assisted Targeting of TGBp2

• Published in: Virology (New York, N.Y.) Vol. 269; no. 1; pp. 113 - 127
• Main Authors: Solovyev, A.G., Stroganova, T.A., Zamyatnin, A.A., Fedorkin, O.N., Schiemann, J., Morozov, S.Yu
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 30.03.2000
• Subjects: Amino Acid Sequence; Biological Transport; Conserved Sequence - genetics; Endoplasmic Reticulum - metabolism; Genes, Viral - genetics; Golgi Apparatus - metabolism; green fluorescent protein; Green Fluorescent Proteins; Hordeivirus; Intracellular Membranes - metabolism; Luminescent Proteins - metabolism; Membrane Proteins - chemistry; Membrane Proteins - genetics; Membrane Proteins - metabolism; Microscopy, Fluorescence; Molecular Sequence Data; Molecular Weight; Mutation - genetics; Nicotiana - cytology; Nicotiana - metabolism; Nicotiana - virology; Organelles - metabolism; Plant Viruses - genetics; Plants, Toxic; Poa semilatent virus; Protein Binding; Protein Sorting Signals - genetics; Protein Sorting Signals - physiology; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; RNA Viruses - genetics; TGBp2 protein; TGBp3 protein; Viral Proteins - chemistry; Viral Proteins - genetics; Viral Proteins - metabolism
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1006/viro.2000.0200

We studied subcellular distribution of green fluorescent protein (GFP)-tagged movement proteins encoded by the second and the third genes of poa semilatent 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 segment(s), whereas the TGBp3 central hydrophilic region was responsible for targeting of GFP-18K. Coexpression of GFP-15K with the intact 18K protein induced drastic changes in the TGBp2 localization: GFP-15K appeared in the cell 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-assisted targeting of TGBp2 to the cell peripheral compartments. This conclusion was further confirmed by similar effects on the PSLV 15K TGBp2 localization induced by TGBp3 proteins of PSLV and potato virus X, which have no detectable sequence similarity to each other.