Dual-colour imaging of membrane protein targeting directed by poa semilatent virus movement protein TGBp3 in plant and mammalian cells.
The movement function of poa semilatent hordeivirus (PSLV) is mediated by the triple gene block (TGB) proteins, of which two, TGBp2 and TGBp3, are membrane proteins. TGBp3 is localized to peripheral bodies in the vicinity of the plasma membrane and is able to re-direct TGBp2 from the endoplasmic reticulum (ER) to the peripheral bodies. For imaging of TGBp3-mediated protein targeting, PSLV TGBp3 tagged with a red fluorescent protein (DsRed) was used. Coexpression of DsRed-TGBp3 with GFP targeted to the ER lumen (ER-GFP) demonstrated that ER-GFP was contained in typical ER structures and peripheral bodies formed by TGBp3 protein, suggesting an ER origin for these bodies. In transient coexpression with viral membrane proteins tagged with GFP, DsRed-TGBp3 directed to the peripheral bodies the homologous TGBp2 protein and two unrelated membrane proteins, the 6 kDa movement protein of beet yellows closterovirus and the putative movement protein encoded by the genome component 4 of faba bean necrotic yellows nanovirus. However, coexpression of TGBp3 with GFP derivatives targeted to the ER membranes by artificial hydrophobic tail sequences suggested that targeting to the ER membranes per se was not sufficient for TGBp3-directed protein trafficking to peripheral bodies. TGBp3-induced targeting of TGBp2 also occurred in mammalian cells, indicating the universal nature of the protein trafficking signals and the cotargeting mechanism.
Zamyatnin
A A
AA
A. N. Belozersky Institute of Physico-Chemical Biology and Department of Virology, Moscow State University, Moscow 119899, Russia1.
Solovyev
A G
AG
A. N. Belozersky Institute of Physico-Chemical Biology and Department of Virology, Moscow State University, Moscow 119899, Russia1.
Sablina
A A
AA
Institute of Carcinogenesis, Cancer Research Center, Moscow 115478, Russia2.
Agranovsky
A A
AA
A. N. Belozersky Institute of Physico-Chemical Biology and Department of Virology, Moscow State University, Moscow 119899, Russia1.
Katul
L
L
Institute of Plant Virology, Microbiology and Biosafety, Federal Biological Research Centre for Agriculture and Forestry, Messeweg 11/12, D-38104 Braunschweig, Germany3.
Vetten
H J
HJ
Institute of Plant Virology, Microbiology and Biosafety, Federal Biological Research Centre for Agriculture and Forestry, Messeweg 11/12, D-38104 Braunschweig, Germany3.
Schiemann
J
J
Institute of Plant Virology, Microbiology and Biosafety, Federal Biological Research Centre for Agriculture and Forestry, Messeweg 11/12, D-38104 Braunschweig, Germany3.
Hinkkanen
A E
AE
Department of Biochemistry and Pharmacy, Åbo Akademi University, 20521 Turku, Finland4.
Lehto
K
K
Department of Biology, University of Turku, 20500 Turku, Finland5.
Morozov
S Yu
SY
A. N. Belozersky Institute of Physico-Chemical Biology and Department of Virology, Moscow State University, Moscow 119899, Russia1.
D000818:Animals; D051242:Caveolin 1; D022461:Caveolins / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D017790:Closterovirus; D003116:Color; D006224:Cricetinae; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009068:Movement; D025682:Nanovirus; D015388:Organelles / Q000378:metabolism; D053830:Plant Viral Movement Proteins; D021382:Protein Sorting Signals / Q000235:genetics / Q000502:physiology; D021381:Protein Transport; D014026:Nicotiana / Q000166:cytology* / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D000097573:Red Fluorescent Protein
D051242:Caveolin 1; D022461:Caveolins; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D008565:Membrane Proteins; D053830:Plant Viral Movement Proteins; D021382:Protein Sorting Signals; D014764:Viral Proteins; D000097573:Red Fluorescent Protein