Distinct sets of tethering complexes, SNARE complexes, and Rab GTPases mediate membrane fusion at the vacuole in Arabidopsis

Takemoto, Kodai; Ebine, Kazuo; Askani, Jana Christin; Krüger, Friedrich; Gonzalez, Zaida Andrés; Ito, Emi; Goh, Tatsuaki; Schumacher, Karin; Nakano, Akihiko; Ueda, Takashi

doi:10.1073/pnas.1717839115

Cited by 113 publications

(122 citation statements)

References 52 publications

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“…the vacuole, PVCs initially interact with RAB5 GTPases that become activated by the GEF protein VPS9; RAB5s then interact with the class C core vacuole/endosome tethering (CORVET) complex composed of the subunits VPS8, VPS3, VPS11, VPS16, VPS18, and VPS33 (11). CORVET permits PVC interaction with the SNAREs VAMP727 and SYP22.…”

Section: Significancementioning

confidence: 99%

Interaction between VPS35 and RABG3f is necessary as a checkpoint to control fusion of late compartments with the vacuole

Rodríguez-Furlán

Domozych

Qian

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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Vacuoles are essential organelles in plants, playing crucial roles, such as cellular material degradation, ion and metabolite storage, and turgor maintenance. Vacuoles receive material via the endocytic, secretory, and autophagic pathways. Membrane fusion is the last step during which prevacuolar compartments (PVCs) and autophagosomes fuse with the vacuole membrane (tonoplast) to deliver cargoes. Protein components of the canonical intracellular fusion machinery that are conserved across organisms, including Arabidopsis thaliana, include complexes, such as soluble N-ethylmaleimide–sensitive factor attachment protein receptors (SNAREs), that catalyze membrane fusion, and homotypic fusion and vacuole protein sorting (HOPS), that serve as adaptors which tether cargo vesicles to target membranes for fusion under the regulation of RAB-GTPases. The mechanisms regulating the recruitment and assembly of tethering complexes are not well-understood, especially the role of RABs in this dynamic regulation. Here, we report the identification of the small synthetic molecule Endosidin17 (ES17), which interferes with synthetic, endocytic, and autophagic traffic by impairing the fusion of late endosome compartments with the tonoplast. Multiple independent target identification techniques revealed that ES17 targets the VPS35 subunit of the retromer tethering complex, preventing its normal interaction with the Arabidopsis RAB7 homolog RABG3f. ES17 interference with VPS35–RABG3f interaction prevents the retromer complex to endosome anchoring, resulting in retention of RABG3f. Using multiple approaches, we show that VPS35–RABG3f–GTP interaction is necessary to trigger downstream events like HOPS complex assembly and fusion of late compartments with the tonoplast. Overall, our results support a role for the interaction of RABG3f–VPS35 as a checkpoint in the control of traffic toward the vacuole.

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Section: Significancementioning

confidence: 99%

Interaction between VPS35 and RABG3f is necessary as a checkpoint to control fusion of late compartments with the vacuole

Rodríguez-Furlán

Domozych

Qian

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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“…By this mechanism, Rab5-to-7 conversion during endosome maturation is linked with the engaging of successive tethers. This mechanism also exists in plants, although recent evidence suggests that the HOPS function in some plant pathways does not depend upon this kind of GTPase switch (Takemoto et al, 2018). In Tetrahymena, our results beg the question of whether there is maturation-linked switching between distinct CORVET complexes like 8C-CC and 8A-CC.…”

Section: Discussionmentioning

confidence: 55%

“…Hence, the potential full diversity of HOPS and CORVET structure and function remains unexplored. Interestingly, recent studies in the Archaeplastida (plants) reveals that the coupling between CORVET and HOPS functions may itself be evolutionarily plastic (Takemoto et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Diversification of CORVET tethers facilitates transport complexity inTetrahymena thermophila

Sparvoli

Zoltner

Field

et al. 2019

Preprint

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In endolysosomal networks, two hetero-hexameric tethers called HOPS and CORVET are found widely throughout eukaryotes. The unicellular ciliate Tetrahymena thermophila possesses elaborate endolysosomal structures, but curiously both it and related protozoa lack the HOPS tether and several other trafficking genes while retaining the related CORVET complex. Tetrahymena encodes multiple paralogs of most CORVET subunits, which assemble into six distinct complexes. Each complex has a unique subunit composition and, significantly, shows unique localization, indicating participation in distinct pathways. One pair of complexes differ by a single subunit (Vps8), but have late endosomal vs. recycling endosome locations. While Vps8 subunits are thus prime determinants for targeting and functional specificity, determinants exist on all subunits except Vps11. This unprecedented expansion and diversification of CORVET provides a potent example of tether flexibility, and illustrates how 'backfilling' following secondary losses of trafficking genes can provide a mechanism for evolution of new pathways. 3

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“…In order to increase the spatial resolution for pH measurements, R-GECO1-P2A-E 2 GFP was fused to the N-terminus of LOW TEMPERATURE INDUCED PROTEIN 6B (LTI6b) or VESICLE TRANSPORT V-SNARE 11 (VTI11). This enabled the targeting of E 2 GFP to the cytosolic side of the plasma membrane (LTI6b; Cutler et al, 2000) or the tonoplast (VTI11; Takemoto et al, 2018), while R-GECO1 remained in the cytosol and the nucleus (Supplemental Figures 3A and 4A). Compared to previous analyses (Figure 5), these indicators reported very similar Ca 2+ and pH response patterns, irrespective of the subcellular localization of E 2 GFP.…”

Section: Resultsmentioning

confidence: 99%

Dual-sensing genetically encoded fluorescent indicators resolve the spatiotemporal coordination of cytosolic abscisic acid and second messenger dynamics in Arabidopsis

Waadt

Köster

Andrés

et al. 2019

Preprint

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Dual-sensing genetically encoded fluorescent indicators resolve the 22 spatiotemporal coordination of cytosolic abscisic acid and second messenger 23 dynamics in Arabidopsis 24 SHORT TITLE 25 Coordination of signaling compound fluxes in roots 26 Rainer Waadt et al. -Coordination of signaling compound fluxes in roots 2 27 ABSTRACT 28 Deciphering signal transduction processes is crucial for understanding 29 how plants sense and respond to environmental changes. Various chemical 30 compounds function as central messengers within deeply intertwined signaling 31 networks. How such compounds act in concert remains to be elucidated. We 32 have developed dual-sensing genetically encoded fluorescent indicators (2-In-1-33 GEFIs) for multiparametric in vivo analyses of the phytohormone abscisic acid 34 (ABA), Ca 2+ , protons (H + ), chloride (anions), the glutathione redox potential (E GSH ) 35 and hydrogen peroxide (H 2 O 2 ). Simultaneous analyses of two signaling 36 compounds in Arabidopsis (Arabidopsis thaliana) roots revealed that ABA 37 treatment and uptake did not trigger rapid cytosolic Ca 2+ or H + fluxes. Glutamate, 38 ATP, Arabidopsis PLANT ELICITOR PEPTIDE (AtPEP1) and glutathione disulfide 39 (GSSG) treatments induced rapid spatiotemporally overlapping cytosolic Ca 2+ , H + 40 and anion fluxes, but except for GSSG only weakly affected the cytosolic redox 41 state. Overall, 2-In-1-GEFIs enable complementary high-resolution in vivo 42 analyses of signaling compound dynamics and facilitate an advanced 43 understanding of the spatiotemporal coordination of signal transduction 44 processes in Arabidopsis. 45 46 198 Supplemental Movie 3). These experiments established the functionality of both 2-In-1-199 GEFIs and revealed that ABA does not trigger rapid cytosolic Ca 2+ or pH changes in 200 roots. 201 Auxin, ATP and glutamate treatments trigger spatiotemporally overlapping fluxes 202 of Ca 2+ , H + and Cl -203 Rainer Waadt et al. -Coordination of signaling compound fluxes in roots 8 Next, we used R-GECO1-GSL-E 2 GFP to simultaneously monitor Ca 2+ , H + and Cl -204 fluxes. Because anions, such as Cl -, quench the fluorescence of E 2 GFP and because its 205 excitation ratiometric pH readout is Clindependent, E 2 GFP provides a means to 206 simultaneously assess cytosolic H + and Clchanges (Bizzarri et al., 2006; Arosio et al., 207 2010). In response to 1 µM IAA, R-GECO1 reported biphasic Ca 2+ signals in the root 208 elongation zone that travelled to neighboring regions. Subsequent 1 mM ATP 209 treatments triggered Ca 2+ signals in the calyptra and meristematic zone that proceeded 210 shootward (Figure 4A left). Interestingly, both Ca 2+ signals coincided with a cytosolic 211 acidification reported by E 2 GFP (Figure 4A middle, Supplemental Movie 4). IAA also 212 induced a Clinflux, indicated by a E 2 GFP fluorescence emission decrease in the entire 213 imaged root, with a maximum decrease in the meristematic zone (Figure 4A right). ATP 214 however induced Clinflux in the upper elongation zone and above, but Clefflux ...

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Distinct sets of tethering complexes, SNARE complexes, and Rab GTPases mediate membrane fusion at the vacuole in Arabidopsis

Cited by 113 publications

References 52 publications

Interaction between VPS35 and RABG3f is necessary as a checkpoint to control fusion of late compartments with the vacuole

Interaction between VPS35 and RABG3f is necessary as a checkpoint to control fusion of late compartments with the vacuole

Diversification of CORVET tethers facilitates transport complexity inTetrahymena thermophila

Dual-sensing genetically encoded fluorescent indicators resolve the spatiotemporal coordination of cytosolic abscisic acid and second messenger dynamics in Arabidopsis

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