Vacuolar and plasma membrane proton pumps collaborate to achieve cytosolic pH homeostasis in yeast.

Martínez-Muñoz, Gloria A.; Kane, Patricia M.

doi:10.1074/jbc.a117.710470

Cited by 87 publications

(50 citation statements)

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“…We used the pH-sensitive fluorophore BCECF, which accumulates in yeast vacuoles. The cells were loaded with BCECF-AM, and the pH was measured fluorometrically according to standard calibration curves (20,24,31). The vacuolar pH was measured in wild-type cells and pfk2⌬ after growing the cells in 2% versus 4% glucose.…”

Section: Stimulation Of Glycolysis Rescues V-atpase Function In Pfk2⌬ Atmentioning

confidence: 99%

“…The V 1 V o complexes reactivate after reassembly, which acidifies the vacuolar lumen and restores membrane gradients and secondary transport systems (20). To determine whether reassembly restored V-ATPase proton transport, we measured the vacuolar pH after readdition of 4% glucose to pfk2⌬.…”

Section: Stimulation Of Glycolysis Rescues V-atpase Function In Pfk2⌬ Atmentioning

confidence: 99%

Section: Vacuolar Hmentioning

confidence: 99%

“…Disassembly is reversed by readdition of glucose to yeast cells (10,21). After reassembly, V 1 V o resumes ATP-driven proton transport, which restores vacuolar and cytosol pH homeostasis (20).Complete glucose depletion causes about 75% of the V 1 V o complexes to disassemble (10, 21); the remaining 25% of the pumps maintain the V-ATPase activity necessary to sustain basal cellular functions. Presumably yeast V 1 V o disassembly preserves energy when glucose, the main energy source, is lim-* This work was supported by National Institutes of Health Grant R01GM086495 (to K. J. P.) and American Heart Association Grant 14PRE19020015 (to C. Y. C.).…”

mentioning

confidence: 99%

“…Disassembly completely inactivates V-ATPase pumps because V 1 without V o cannot hydrolyze ATP, and V o without V 1 cannot transport protons (2). Consequently, protons cannot not be redistributed from the cytosol into the vacuole, which alkalinizes the vacuolar lumen and acidifies the cytosol (19,20). Disassembly is reversed by readdition of glucose to yeast cells (10,21).…”

mentioning

confidence: 99%

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Regulation of Vacuolar H+-ATPase (V-ATPase) Reassembly by Glycolysis Flow in 6-Phosphofructo-1-kinase (PFK-1)-deficient Yeast Cells

Chan¹,

Dominguez

Parra

2016

Journal of Biological Chemistry

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Vacuolar H ϩ -ATPase (V-ATPase) 3 is a highly conserved ATP-driven proton pump distributed throughout the endomembrane system. Intracellular V-ATPase generates and maintains the acidic organelle luminal pH necessary for membrane trafficking, protein sorting and processing, and zymogen activation (1-5). V-ATPase is also present at the plasma membrane of cells specialized for active proton transport. Kidney intercalated cells (6), bone osteoclasts (7), and epididymis clear cells (6) express V-ATPases at the plasma membrane, which are necessary for systemic acid-base balance, bone resorption, and sperm maturation, respectively.V-ATPase consists of two multisubunit domains, V 1 and V o , that are responsible for ATP hydrolysis and proton transport, respectively (8, 9). Peripheral subunits form V 1 , the catalytic domain attached to the cytosolic side of the membrane. V 1 is bound to V o , the integral membrane domain that forms the path for proton transport. During catalysis, ATP hydrolysis within V 1 drives active transport of protons from the cytosol to the other side of the membrane via rotation of a proteolipid ring structure in V o . Detachment of V 1 from V o is an important mechanism that inhibits V-ATPase proton transport (3, 10, 11). Disassembly and reassembly of V 1 V o has been observed in yeast, insects, and mammalian cells (3,(11)(12)(13). However, the cellular mechanisms governing V-ATPase regulation by reversible disassembly are not well understood. In yeast, V-ATPase reversible disassembly is intertwined with cytosol pH changes (14, 15) the RAS/cAMP/PKA pathway (16), and glycolysis (3).Disassembly of the yeast V 1 V o complex occurs when glucose is not available. Upon glucose depletion, V 1 subunit C is released into the cytoplasm, causing the rest of the V 1 domain to dissociate from V o (10,17,18). Disassembly completely inactivates V-ATPase pumps because V 1 without V o cannot hydrolyze ATP, and V o without V 1 cannot transport protons (2). Consequently, protons cannot not be redistributed from the cytosol into the vacuole, which alkalinizes the vacuolar lumen and acidifies the cytosol (19,20). Disassembly is reversed by readdition of glucose to yeast cells (10,21). After reassembly, V 1 V o resumes ATP-driven proton transport, which restores vacuolar and cytosol pH homeostasis (20).Complete glucose depletion causes about 75% of the V 1 V o complexes to disassemble (10, 21); the remaining 25% of the pumps maintain the V-ATPase activity necessary to sustain basal cellular functions. Presumably yeast V 1 V o disassembly preserves energy when glucose, the main energy source, is lim-* This work was supported by National Institutes of Health Grant R01GM086495 (to K. J. P.) and American Heart Association Grant 14PRE19020015 (to C. Y. C.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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Section: Stimulation Of Glycolysis Rescues V-atpase Function In Pfk2⌬ Atmentioning

confidence: 99%

Section: Stimulation Of Glycolysis Rescues V-atpase Function In Pfk2⌬ Atmentioning

confidence: 99%

Section: Vacuolar Hmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Regulation of Vacuolar H+-ATPase (V-ATPase) Reassembly by Glycolysis Flow in 6-Phosphofructo-1-kinase (PFK-1)-deficient Yeast Cells

Chan¹,

Dominguez

Parra

2016

Journal of Biological Chemistry

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Sterol uptake by the NPC system in eukaryotes: a Saccharomyces cerevisiae perspective

et al. 2021

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Sterols are an essential component of membranes in all eukaryotic cells and the precursor of multiple indispensable cellular metabolites. After endocytotic uptake, sterols are integrated into the lysosomal membrane by the Niemann-Pick type C (NPC) system before redistribution to other membranes. The process is driven by two proteins that, together, compose the NPC system: the lysosomal sterol shuttle protein NPC2 and the membrane protein NPC1 (named NCR1 in fungi), which integrates sterols into the lysosomal membrane. The Saccharomyces cerevisiae NPC system provides a compelling model to study the molecular mechanism of sterol integration into membranes and sterol homeostasis. This review summarizes recent advances in the field, and by interpreting available structural data, we propose a unifying conceptual model for sterol loading, transfer and transport by NPC proteins.

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Visible‐Light‐Activated Molecular Machines Kill Fungi by Necrosis Following Mitochondrial Dysfunction and Calcium Overload

et al. 2023

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Invasive fungal infections are a growing public health threat. As fungi become increasingly resistant to existing drugs, new antifungals are urgently needed. Here, it is reported that 405‐nm‐visible‐light‐activated synthetic molecular machines (MMs) eliminate planktonic and biofilm fungal populations more effectively than conventional antifungals without resistance development. Mechanism‐of‐action studies show that MMs bind to fungal mitochondrial phospholipids. Upon visible light activation, rapid unidirectional drilling of MMs at ≈3 million cycles per second (MHz) results in mitochondrial dysfunction, calcium overload, and ultimately necrosis. Besides their direct antifungal effect, MMs synergize with conventional antifungals by impairing the activity of energy‐dependent efflux pumps. Finally, MMs potentiate standard antifungals both in vivo and in an ex vivo porcine model of onychomycosis, reducing the fungal burden associated with infection.

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Vacuolar and plasma membrane proton pumps collaborate to achieve cytosolic pH homeostasis in yeast.

Cited by 87 publications

References 68 publications

Regulation of Vacuolar H+-ATPase (V-ATPase) Reassembly by Glycolysis Flow in 6-Phosphofructo-1-kinase (PFK-1)-deficient Yeast Cells

Regulation of Vacuolar H+-ATPase (V-ATPase) Reassembly by Glycolysis Flow in 6-Phosphofructo-1-kinase (PFK-1)-deficient Yeast Cells

Sterol uptake by the NPC system in eukaryotes: a Saccharomyces cerevisiae perspective

Visible‐Light‐Activated Molecular Machines Kill Fungi by Necrosis Following Mitochondrial Dysfunction and Calcium Overload

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