Direct interaction of the Golgi V-ATPase a-subunit isoform with PI(4)P drives localization of Golgi V-ATPases in yeast

Banerjee, Subhrajit; Kane, Patricia M.

doi:10.1091/mbc.e17-05-0316

Cited by 41 publications

(71 citation statements)

References 82 publications

(122 reference statements)

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“…The phosphatidylinositol lipid PI(4)P is enriched in the Golgi membranes of yeast but not in the vacuolar membranes, and has been shown to interact specifically with Stv1p but not Vph1p (15). PI(4)P is required for appropriate localization of Stv1-containing V-ATPases, and reduction in intracellular PI(4)P levels has also been shown to impair V-ATPase activity (15). In ATPase assays of the purified protein, the addition of PI(4)P did not significantly affect the activity of Vph1-…”

Section: Resultsmentioning

confidence: 99%

“…The N-terminal domain of Stv1p contains the W 83 KY sequence, which is necessary and sufficient for targeting Stv1-V 1 V O to the Golgi apparatus (14). K84 of this sequence has been shown to interact specifically with the phosphatidylinositol lipid PI(4)P, which is required for efficient localization and activity of the Golgi V-ATPase (15). Interestingly, no density was observed in the Stv1-V O map for the region corresponding to H78 to L111, which contains the Golgi targeting sequence, suggesting that this region is flexible (SI Appendix, Fig.…”

Section: Overall Structures Of Stv1-v O and Vph1-v Omentioning

confidence: 99%

“…Vph1p, the major isoform, is found in the vacuole, while Stv1p is found in the Golgi apparatus and endosomes (13). The soluble N-terminal domain of subunit a is required for targeting V-ATPase to its appropriate cellular location (13)(14)(15). The membrane-embedded C-terminal domain contains two half-channels for proton translocation and, together with the c 8 c′c″-ring (c-ring), carries out transmembrane proton transport by a mechanism similar to that of the F-type ATP synthase (1,8,(16)(17)(18).…”

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confidence: 99%

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Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae

Vasanthakumar

Bueler

et al. 2019

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

112

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Proton-translocating vacuolar-type ATPases (V-ATPases) are necessary for numerous processes in eukaryotic cells, including receptor-mediated endocytosis, protein maturation, and lysosomal acidification. In mammals, V-ATPase subunit isoforms are differentially targeted to various intracellular compartments or tissues, but how these subunit isoforms influence enzyme activity is not clear. In the yeast Saccharomyces cerevisiae, isoform diversity is limited to two different versions of the proton-translocating subunit a: Vph1p, which is targeted to the vacuole, and Stv1p, which is targeted to the Golgi apparatus and endosomes. We show that purified V-ATPase complexes containing Vph1p have higher ATPase activity than complexes containing Stv1p and that the relative difference in activity depends on the presence of lipids. We also show that VO complexes containing Stv1p could be readily purified without attached V1 regions. We used this effect to determine structures of the membrane-embedded VO region with Stv1p at 3.1-Å resolution, which we compare with a structure of the VO region with Vph1p that we determine to 3.2-Å resolution. These maps reveal differences in the surface charge near the cytoplasmic proton half-channel. Both maps also show the presence of bound lipids, as well as regularly spaced densities that may correspond to ergosterol or bound detergent, around the c-ring.

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Section: Resultsmentioning

confidence: 99%

Section: Overall Structures Of Stv1-v O and Vph1-v Omentioning

confidence: 99%

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confidence: 99%

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Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae

Vasanthakumar

Bueler

et al. 2019

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

112

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“…In yeast, all the subunits of the V-ATPase have only one isoform, except the "a" subunit of the V 0 domain which exists as two isoforms. Vph1p mainly localizes in the vacuole, while Stv1p is found in V-ATPase complexes of the Golgi apparatus and of endosomes [37][38][39][40] . Using the new Golgi pH probe, we show that deletion of STV1 only slightly increases the www.nature.com/scientificreports www.nature.com/scientificreports/ Golgi pH (Fig.…”

Section: Resultsmentioning

confidence: 99%

A new pH sensor localized in the Golgi apparatus of Saccharomyces cerevisiae reveals unexpected roles of Vph1p and Stv1p isoforms

Deschamps

Colinet

Zimmermannová

et al. 2020

Sci Rep

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The gradual acidification of the secretory pathway is conserved and extremely important for eukaryotic cells, but until now there was no pH sensor available to monitor the pH of the early Golgi apparatus in Saccharomyces cerevisiae. therefore, we developed a pHluorin-based sensor for in vivo measurements in the lumen of the Golgi. By using this new tool we show that the cis-and medial-Golgi pH is equal to 6.6-6.7 in wild type cells during exponential phase. As expected, V-ATPase inactivation results in a near neutral Golgi pH. We also uncover that surprisingly Vph1p isoform of the V-ATPase is prevalent to Stv1p for Golgi acidification. Additionally, we observe that during changes of the cytosolic pH, the Golgi pH is kept relatively stable, mainly thanks to the V-ATPase. Eventually, this new probe will allow to better understand the mechanisms involved in the acidification and the pH control within the secretory pathway.

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“…This interaction stabilizes the assembly of V1-Vo complex to form the active V-ATPase (Banerjee et al, 2019, Li et al, 2014. In the Golgi, Vph1 is replaced by Stv1 and interacts with the compartment rich lipid PI4P instead of PI(3,5)P2, which is only made on late endosomes and lysosomes (Banerjee and Kane, 2017). In both instances, it is clear that specific phosphoinositides are essential for V-ATPase function.…”

Section: Introductionmentioning

confidence: 99%

Reciprocal regulation of vacuolar calcium transport and V-ATPase activity, and the effects of Phosphatidylinositol 3,5-bisphosphate

Miner

Rivera‐Kohr

Zhang

et al. 2020

Preprint

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StatementHere we show that Ca 2+ and H + transport across the vacuole membrane is reciprocally regulated and that it is linked to the production of Phosphatidylinositol 3,5-bisphoshpate. ABSTRACTYeast vacuoles are acidified by the V-ATPase, a protein complex comprised of the membrane embedded VO complex and the soluble cytoplasmic V1 complex. The assembly of the V1-VO holoenzyme is required for the transfer of H + into the vacuole lumen for acidification. The assembly of the V1-VO holoenzyme is stabilized by the lipid phosphatidylinositol 3,5-bisphospate (PI(3,5)P2) made by the PI3P 5-kinase Fab1/PIKfyve. The absence of PI(3,5)P2 leads to the dissociation of the V1 complex from the membrane. Separately, PI(3,5)P2 has been shown to modulate Ca 2+ transport across the vacuole membrane during fission and fusion. Here we examined whether the regulation of H + and Ca 2+ by PI(3,5)P2 are interdependent. We show that modulating extraluminal Ca 2+ concentrations inhibit V-ATPase activity. As extraluminal CaCl2 levels are raised, the activity of H + pumping is reduced. Conversely, chelating free Ca 2+ with EGTA stimulated vacuole acidification. Not only did Ca 2+ levels affect H + translocation, we also show that blocking V-ATPase activity inhibited Ca 2+ transport into the vacuole lumen. Together, these data illustrate that Ca 2+ transport and V-ATPase regulation are interconnected through the modulation of vacuolar lipid profiles. Abbreviations: AO, acridine orange; DAG, diacylglycerol; FCCP, Carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone; PA, phosphatidic acid; PI, phosphatidylinositol; PI3P, phosphatidylinositol 3-phosphate; PI(3,5)P2, phosphatidylinositol 3,5-bisphosphate; PKC, protein kinase C; PLC, phospholipase C; SNARE, soluble N-ethylmaleimide-sensitive factor attachment protein receptor; TRP, transient receptor potential; YPD, yeast extract, peptone, dextrose. Miner et al. -Reciprocal regulation of V-ATPase and Ca 2+ transport

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Direct interaction of the Golgi V-ATPase a-subunit isoform with PI(4)P drives localization of Golgi V-ATPases in yeast

Cited by 41 publications

References 82 publications

Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae

Structural comparison of the vacuolar and Golgi V-ATPases from Saccharomyces cerevisiae

A new pH sensor localized in the Golgi apparatus of Saccharomyces cerevisiae reveals unexpected roles of Vph1p and Stv1p isoforms

Reciprocal regulation of vacuolar calcium transport and V-ATPase activity, and the effects of Phosphatidylinositol 3,5-bisphosphate

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