AtRabF2b (Ara7) acts on the vacuolar trafficking pathway in tobacco leaf epidermal cells.
Rab GTPases are universal key regulators of intracellular secretory trafficking events. In particular, Rab 5 homologues have been implicated in endocytic events and in the vacuolar pathway. In this study, we investigate the location and function of a member of this family, AtRabF2b (Ara7) in tobacco (Nicotiana tabacum) leaf epidermal cells using a live cell imaging approach. Fluorescent-tagged AtRabF2b[wt] localized to the prevacuolar compartment and Golgi apparatus, as determined by coexpression studies with fluorescent markers for these compartments. Mutations that impair AtRabF2b function also alter the subcellular location of the GTPase. In addition, coexpression studies of the protein with the vacuole-targeted aleurain-green fluorescent protein (GFP) and rescue experiments with wild-type AtRabF2b indicate that the dominant-negative mutant of AtRabF2b causes the vacuolar marker to be secreted to the apoplast. Our results indicate a clear role of AtRabF2b in the vacuolar trafficking pathway.
Kotzer
Amanda M
AM
Research School of Biological and Molecular Sciences, Oxford Brookes University, Gipsy Lane, Oxford, OX3 0BP, UK.
Brandizzi
Federica
F
Neumann
Ulla
U
Paris
Nadine
N
Moore
Ian
I
Hawes
Chris
C
D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D006056:Golgi Apparatus / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008954:Models, Biological; D009154:Mutation; D018515:Plant Leaves / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014026:Nicotiana / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism*; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
D015415:Biomarkers; D010940:Plant Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D020691:rab GTP-Binding Proteins