The factors and mechanisms involved in vacuolar transport in plants, and in particular those directing vesicles to their target endomembrane compartment, remain largely unknown. To identify components of the vacuolar trafficking machinery, we searched for Arabidopsis modified transport to the vacuole (mtv) mutants that abnormally secrete the synthetic vacuolar cargo VAC2. We report here on the identification of 17 mtv mutations, corresponding to mutant alleles of MTV2/VSR4, MTV3/PTEN2A MTV7/EREL1, MTV8/ARFC1, MTV9/PUF2, MTV10/VPS3, MTV11/VPS15, MTV12/GRV2, MTV14/GFS10, MTV15/BET11, MTV16/VPS51, MTV17/VPS54, and MTV18/VSR1 Eight of the MTV proteins localize at the interface between the trans-Golgi network (TGN) and the multivesicular bodies (MVBs), supporting that the trafficking step between these compartments is essential for segregating vacuolar proteins from those destined for secretion. Importantly, the GARP tethering complex subunits MTV16/VPS51 and MTV17/VPS54 were found at endoplasmic reticulum (ER)- and microtubule-associated compartments (EMACs). Moreover, MTV16/VPS51 interacts with the motor domain of kinesins, suggesting that, in addition to tethering vesicles, the GARP complex may regulate the motors that transport them. Our findings unveil a previously uncharacterized compartment of the plant vacuolar trafficking pathway and support a role for microtubules and kinesins in GARP-dependent transport of soluble vacuolar cargo in plants.

Central European Institute of Technology Mendel University in Brno CZ 61300 Brno Czech Republic

Centro Nacional de Biotecnología Consejo Superior de Investigaciones Científicas Cantoblanco E 28049 Madrid Spain

Centro Nacional de Biotecnología Consejo Superior de Investigaciones Científicas Cantoblanco E 28049 Madrid Spain;

Department of Biosciences Durham University Durham DH1 3LE United Kingdom

Department of Plant Physiology University of Potsdam 14476 Potsdam Germany

Key Laboratory of Horticultural Plant Biology College of Horticulture and Forestry Sciences Huazhong Agricultural University Wuhan 430070 China

School of Life Sciences Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology The Chinese University of Hong Kong Shatin New Territories Hong Kong China

State Key Laboratory of Subtropical Silviculture Zhejiang A and F University Hangzhou 311300 China

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Proc Natl Acad Sci U S A. 2020 May 19;117(20):10623-10624. doi: 10.1073/pnas.2006766117. PubMed

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