Amino Acid Availability Modulates Vacuolar H+-ATPase Assembly
- PMID: 26378229
- PMCID: PMC4646367
- DOI: 10.1074/jbc.M115.659128
Amino Acid Availability Modulates Vacuolar H+-ATPase Assembly
Abstract
The vacuolar H(+)-ATPase (V-ATPase) is an ATP-dependent proton pump composed of a peripheral ATPase domain (V1) and a membrane-integral proton-translocating domain (V0) and is involved in many normal and disease processes. An important mechanism of regulating V-ATPase activity is reversible assembly of the V1 and V0 domains. Increased assembly in mammalian cells occurs under various conditions and has been shown to involve PI3K. The V-ATPase is necessary for amino acid-induced activation of mechanistic target of rapamycin complex 1 (mTORC1), which is important in controlling cell growth in response to nutrient availability and growth signals. The V-ATPase undergoes amino acid-dependent interactions with the Ragulator complex, which is involved in recruitment of mTORC1 to the lysosomal membrane during amino acid sensing. We hypothesized that changes in the V-ATPase/Ragulator interaction might involve amino acid-dependent changes in V-ATPase assembly. To test this, we measured V-ATPase assembly by cell fractionation in HEK293T cells treated with and without amino acids. V-ATPase assembly increases upon amino acid starvation, and this effect is reversed upon readdition of amino acids. Lysosomes from amino acid-starved cells possess greater V-ATPase-dependent proton transport, indicating that assembled pumps are catalytically active. Amino acid-dependent changes in both V-ATPase assembly and activity are independent of PI3K and mTORC1 activity, indicating the involvement of signaling pathways distinct from those implicated previously in controlling assembly. By contrast, lysosomal neutralization blocks the amino acid-dependent change in assembly and reactivation of mTORC1 after amino acid starvation. These results identify an important new stimulus for controlling V-ATPase assembly.
Keywords: amino acid; lysosomal acidification; mechanistic target of rapamycin (mTOR); nutrient sensing; proton transport; regulated assembly; vacuolar ATPase.
© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.
Figures
Similar articles
-
Regulated assembly of vacuolar ATPase is increased during cluster disruption-induced maturation of dendritic cells through a phosphatidylinositol 3-kinase/mTOR-dependent pathway.J Biol Chem. 2014 Jan 17;289(3):1355-63. doi: 10.1074/jbc.M113.524561. Epub 2013 Nov 22. J Biol Chem. 2014. PMID: 24273170 Free PMC article.
-
mTORC1 senses lysosomal amino acids through an inside-out mechanism that requires the vacuolar H(+)-ATPase.Science. 2011 Nov 4;334(6056):678-83. doi: 10.1126/science.1207056. Science. 2011. PMID: 22053050 Free PMC article.
-
AKT Ser/Thr kinase increases V-ATPase-dependent lysosomal acidification in response to amino acid starvation in mammalian cells.J Biol Chem. 2020 Jul 10;295(28):9433-9444. doi: 10.1074/jbc.RA120.013223. Epub 2020 May 14. J Biol Chem. 2020. PMID: 32409581 Free PMC article.
-
The emerging roles of vacuolar-type ATPase-dependent Lysosomal acidification in neurodegenerative diseases.Transl Neurodegener. 2020 May 11;9(1):17. doi: 10.1186/s40035-020-00196-0. Transl Neurodegener. 2020. PMID: 32393395 Free PMC article. Review.
-
Regulation of V-ATPase assembly and function of V-ATPases in tumor cell invasiveness.Biochim Biophys Acta. 2016 Aug;1857(8):1213-1218. doi: 10.1016/j.bbabio.2016.02.010. Epub 2016 Feb 22. Biochim Biophys Acta. 2016. PMID: 26906430 Free PMC article. Review.
Cited by
-
ATP6V1B1 regulates ovarian cancer progression and cisplatin sensitivity through the mTOR/autophagy pathway.Mol Cell Biochem. 2024 May 12. doi: 10.1007/s11010-024-05025-w. Online ahead of print. Mol Cell Biochem. 2024. PMID: 38735913
-
Molecular mechanism of Oxr1p mediated disassembly of yeast V-ATPase.EMBO Rep. 2024 May;25(5):2323-2347. doi: 10.1038/s44319-024-00126-5. Epub 2024 Apr 2. EMBO Rep. 2024. PMID: 38565737 Free PMC article.
-
TM4SF19-mediated control of lysosomal activity in macrophages contributes to obesity-induced inflammation and metabolic dysfunction.Nat Commun. 2024 Mar 30;15(1):2779. doi: 10.1038/s41467-024-47108-8. Nat Commun. 2024. PMID: 38555350 Free PMC article.
-
Ubiquitin ligase subunit FBXO9 inhibits V-ATPase assembly and impedes lung cancer metastasis.Exp Hematol Oncol. 2024 Mar 14;13(1):32. doi: 10.1186/s40164-024-00497-4. Exp Hematol Oncol. 2024. PMID: 38486234 Free PMC article.
-
Organelle proteomic profiling reveals lysosomal heterogeneity in association with longevity.Elife. 2024 Jan 19;13:e85214. doi: 10.7554/eLife.85214. Elife. 2024. PMID: 38240316 Free PMC article.
References
-
- Forgac M. (2007) Vacuolar ATPases: rotary proton pumps in physiology and pathophysiology. Nat. Rev. Mol. Cell Biol. 8, 917–929 - PubMed
-
- Kane P. M. (1995) Disassembly and reassembly of the yeast vacuolar H+-ATPase in vivo. J. Biol. Chem. 270, 17025–17032 - PubMed
-
- Sumner J. P., Dow J. A., Earley F. G., Klein U., Jäger D., and Wieczorek H. (1995) Regulation of plasma membrane V-ATPase activity by dissociation of peripheral subunits. J. Biol. Chem. 270, 5649–5653 - PubMed
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous