Abstract
Members of the Rab family of small GTPases play important roles in membrane trafficking along the exocytic and endocytic pathways, their function being dependent on their localization. Here, we show the vital roles of the Oryza sativa Rab11 (OsRab11) in intracellular trafficking using a dominant-negative mutant approach based on a protoplast-trafficking assay. GTP-binding and GTP hydrolysis assays revealed that OsRab11 is a small GTP-binding protein complementing △ypt32/ypt31. Green fluorescent protein (GFP)-OsRab11 has been localized to both the trans-Golgi-network (TGN) and the endosomes/prevacuolar compartments (PVC) in Arabidopsis protoplasts. The protoplast transformation with the dominant-negative mutant OsRab11(S28N) revealed that the trafficking of plasma membrane marker proteins [H+-ATPase-GFP and Ca2+-ATPase8-GFP (ACA8-GFP)] and central vacuole marker proteins [Arabidopsis aleurain-like protein (AALP-GFP) and sporamin (Spo-GFP)] was inhibited. Moreover, overexpression of Arabidopsis AtRha1 did not recover the trafficking inhibition of marker proteins from the central vacuole. These results strongly indicate that OsRab11 localizes to either the TGN or the PVC, and plays a significant role in the intracellular trafficking from the TGN to the PM and/or to the PVC in planta.
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Acknowledgments
This work was supported by a grant from the Next-Generation BioGreen 21 Program (SSAC, Grant#: PJ0111062015) and by “Cooperative Research Program for Agriculture Science & Technology development (Project No. PJ009759)” Rural Development Administration, Republic of Korea and by the National Research Foundation of Korea (NRF) Grant funded by Korean Government (NRF-2014R1A1A4A01007558).
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Yun, H.R., Rim, Y.G. & Heo, J.B. Rice small GTPase Rab11 is required for intracellular trafficking from the trans-Golgi-network to the plasma membrane and/or prevacuolar compartments. Appl Biol Chem 59, 163–171 (2016). https://doi.org/10.1007/s13765-015-0143-6
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DOI: https://doi.org/10.1007/s13765-015-0143-6