CoChat
Sign Up
Cited by 424

The DOI matches its Crossref record; 6 of the 6 references it cites were found in a registry or an index.

Autophagosome–lysosome fusion is independent of V-ATPase-mediated acidification

Mauvezin, Caroline, Nagy, Péter, Juhász, Gábor, Neufeld, Thomas P.·2015·Nature Communications
DOI 10.1038/ncomms8007PMC PMC4428688
Open this paper in CoChat — free account

Abstract

The ATP-dependent proton pump V-ATPase ensures low intralysosomal pH, which is essential for lysosomal hydrolase activity. Based on studies with the V-ATPase inhibitor BafilomycinA1, lysosomal acidification is also thought to be required for fusion with incoming vesicles from the autophagic and endocytic pathways. Here we show that loss of V-ATPase subunits in the Drosophila fat body causes an accumulation of non-functional lysosomes, leading to a block in autophagic flux. However, V-ATPase-deficient lysosomes remain competent to fuse with autophagosomes and endosomes, resulting in a time-dependent formation of giant autolysosomes. In contrast, BafilomycinA1 prevents autophagosome–lysosome fusion in these cells, and this defect is phenocopied by depletion of the Ca 2+ pump SERCA, a secondary target of this drug. Moreover, activation of SERCA promotes fusion in a BafilomycinA1-sensitive manner. Collectively, our results indicate that lysosomal acidification is not a prerequisite for fusion, and that BafilomycinA1 inhibits fusion independent of its effect on lysosomal pH.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1038/ncomms8007

Cites

6 of 6 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Flagged first, then by citations

Show all 6 references

Research works printed but not linked: 64

  1. Beau, I., Mehrpour, M. & Codogno, P. Autophagosomes and human diseases. Int. J. Biochem. Cell Biol. 43, 460–464 (2011). · DOI
  2. Mauvezin, C., Ayala, C., Braden, C. R., Kim, J. & Neufeld, T. P. Assays to monitor autophagy in Drosophila. Methods 68, 134–139 (2014). · DOI
  3. Shibutani, S. T. & Yoshimori, T. A current perspective of autophagosome biogenesis. Cell Res. 24, 58–68 (2014). · DOI
  4. Dechant, R. et al. Cytosolic pH is a second messenger for glucose and regulates the PKA pathway through V-ATPase. EMBO J. 29, 2515–2526 (2010). · DOI
  5. Alzamora, R. et al. PKA regulates vacuolar H+-ATPase localization and activity via direct phosphorylation of the a subunit in kidney cells. J. Biol. Chem. 285, 24676–24685 (2010). · DOI
  6. Marshansky, V. & Futai, M. The V-type H+-ATPase in vesicular trafficking: targeting, regulation and function. Curr. Opin. Cell Biol. 20, 415–426 (2008). · DOI
  7. Rath, S., Liebl, J., Furst, R., Vollmar, A. M. & Zahler, S. Regulation of endothelial signaling and migration by v-ATPase. Angiogenesis 17, 587–601 (2014). · DOI
  8. Hinton, A., Bond, S. & Forgac, M. V-ATPase functions in normal and disease processes. Pflugers Arch. 457, 589–598 (2009). · DOI
  9. Zhang, J., Feng, Y. & Forgac, M. Proton conduction and bafilomycin binding by the V0 domain of the coated vesicle V-ATPase. J. Biol. Chem. 269, 23518–23523 (1994). · DOI
  10. Yoshimori, T., Yamamoto, A., Moriyama, Y., Futai, M. & Tashiro, Y. Bafilomycin A1, a specific inhibitor of vacuolar-type H(+)-ATPase, inhibits acidification and protein degradation in lysosomes of cultured cells. J. Biol. Chem. 266, 17707–17712 (1991). · DOI
  11. Wang, D. et al. Ca2+-Calmodulin regulates SNARE assembly and spontaneous neurotransmitter release via v-ATPase subunit V0a1. J. Cell Biol. 205, 21–31 (2014). · DOI
  12. Marshansky, V., Rubinstein, J. L. & Gruber, G. Eukaryotic V-ATPase: novel structural findings and functional insights. Biochim. Biophys. Acta. 1837, 857–879 (2014). · DOI
  13. Forgac, M. Vacuolar ATPases: rotary proton pumps in physiology and pathophysiology. Nat. Rev. Mol. Cell Biol. 8, 917–929 (2007). · DOI
  14. Scott, R. C., Schuldiner, O. & Neufeld, T. P. Role and regulation of starvation-induced autophagy in the Drosophila fat body. Dev. Cell 7, 167–178 (2004). · DOI
  15. Rusten, T. E. et al. Programmed autophagy in the Drosophila fat body is induced by ecdysone through regulation of the PI3K pathway. Dev. Cell 7, 179–192 (2004). · DOI
  16. Lindmo, K. et al. The PI 3-kinase regulator Vps15 is required for autophagic clearance of protein aggregates. Autophagy 4, 500–506 (2008). · DOI
  17. Chang, Y. Y. & Neufeld, T. P. An Atg1/Atg13 complex with multiple roles in TOR-mediated autophagy regulation. Mol. Biol. Cell 20, 2004–2014 (2009). · DOI
  18. Nezis, I. P. et al. Ref(2)P, the Drosophila melanogaster homologue of mammalian p62, is required for the formation of protein aggregates in adult brain. J. Cell Biol. 180, 1065–1071 (2008). · DOI · Paper page
  19. Pircs, K. et al. Advantages and limitations of different p62-based assays for estimating autophagic activity in Drosophila. PLoS ONE 7, e44214 (2012). · DOI
  20. Kovacs, A. L., Reith, A. & Seglen, P. O. Accumulation of autophagosomes after inhibition of hepatocytic protein degradation by vinblastine, leupeptin or a lysosomotropic amine. Exp. Cell Res. 137, 191–201 (1982). · DOI · Paper page
  21. Klionsky, D. J., Elazar, Z., Seglen, P. O. & Rubinsztein, D. C. Does bafilomycin A1 block the fusion of autophagosomes with lysosomes? Autophagy 4, 849–850 (2008). · DOI · Paper page
  22. Drose, S. & Altendorf, K. Bafilomycins and concanamycins as inhibitors of V-ATPases and P-ATPases. J. Exp. Biol. 200, 1–8 (1997). · DOI
  23. Takats, S. et al. Autophagosomal Syntaxin17-dependent lysosomal degradation maintains neuronal function in Drosophila. J. Cell Biol. 201, 531–539 (2013). · DOI
  24. Itakura, E., Kishi-Itakura, C. & Mizushima, N. The hairpin-type tail-anchored SNARE syntaxin 17 targets to autophagosomes for fusion with endosomes/lysosomes. Cell 151, 1256–1269 (2012). · DOI
  25. Baba, M., Osumi, M. & Ohsumi, Y. Analysis of the membrane structures involved in autophagy in yeast by freeze-replica method. Cell Struct. Funct. 20, 465–471 (1995). · DOI · Paper page
  26. Takeshige, K., Baba, M., Tsuboi, S., Noda, T. & Ohsumi, Y. Autophagy in yeast demonstrated with proteinase-deficient mutants and conditions for its induction. J. Cell Biol. 119, 301–311 (1992). · DOI
  27. Bowman, B. J. & Bowman, E. J. Mutations in subunit C of the vacuolar ATPase confer resistance to bafilomycin and identify a conserved antibiotic binding site. J. Biol. Chem. 277, 3965–3972 (2002). · DOI
  28. Jahreiss, L., Menzies, F. M. & Rubinsztein, D. C. The itinerary of autophagosomes: from peripheral formation to kiss-and-run fusion with lysosomes. Traffic. 9, 574–587 (2008). · DOI
  29. Kim, J. & Neufeld, T. P. Dietary sugar promotes systemic TOR activation in Drosophila through AKH-dependent selective secretion of Dilp3. Nat. Commun doi:10.1038/ncomms7846 (2015). · DOI
  30. Bowman, E. J., Siebers, A. & Altendorf, K. Bafilomycins: a class of inhibitors of membrane ATPases from microorganisms, animal cells, and plant cells. Proc. Natl Acad. Sci. USA 85, 7972–7976 (1988). · DOI
  31. Gunteski-Hamblin, A. M., Greeb, J. & Shull, G. E. A novel Ca2+ pump expressed in brain, kidney, and stomach is encoded by an alternative transcript of the slow-twitch muscle sarcoplasmic reticulum Ca-ATPase gene. J. Biol. Chem. 263, 15032–15040 (1988). · DOI
  32. Lytton, J., Westlin, M. & Hanley, M. R. Thapsigargin inhibits the sarcoplasmic or endoplasmic reticulum Ca-ATPase family of calcium pumps. J. Biol. Chem. 266, 17067–17071 (1991). · DOI
  33. Ganley, I. G., Wong, P. M., Gammoh, N. & Jiang, X. Distinct autophagosomal-lysosomal fusion mechanism revealed by thapsigargin-induced autophagy arrest. Mol. Cell 42, 731–743 (2011). · DOI
  34. Bi, J. et al. Seipin promotes adipose tissue fat storage through the ER Ca(2)(+)-ATPase SERCA. Cell Metab. 19, 861–871 (2014). · DOI
  35. Sagne, C. & Gasnier, B. Molecular physiology and pathophysiology of lysosomal membrane transporters. J. Inherit. Metab. Dis. 31, 258–266 (2008). · DOI
  36. Christensen, K. A., Myers, J. T. & Swanson, J. A. pH-dependent regulation of lysosomal calcium in macrophages. J. Cell Sci. 115, 599–607 (2002). · DOI
  37. Strasser, B., Iwaszkiewicz, J., Michielin, O. & Mayer, A. The V-ATPase proteolipid cylinder promotes the lipid-mixing stage of SNARE-dependent fusion of yeast vacuoles. EMBO J. 30, 4126–4141 (2011). · DOI
  38. Williamson, W. R., Wang, D., Haberman, A. S. & Hiesinger, P. R. A dual function of V0-ATPase a1 provides an endolysosomal degradation mechanism in Drosophila melanogaster photoreceptors. J. Cell Biol. 189, 885–899 (2010). · DOI
  39. Baars, T. L., Petri, S., Peters, C. & Mayer, A. Role of the V-ATPase in regulation of the vacuolar fission-fusion equilibrium. Mol. Biol. Cell. 18, 3873–3882 (2007). · DOI
  40. Coonrod, E. M. et al. Homotypic vacuole fusion in yeast requires organelle acidification and not the V-ATPase membrane domain. Dev. Cell 27, 462–468 (2013). · DOI
  41. Poea-Guyon, S. et al. The V-ATPase membrane domain is a sensor of granular pH that controls the exocytotic machinery. J. Cell Biol. 203, 283–298 (2013). · DOI
  42. Kawai, A., Uchiyama, H., Takano, S., Nakamura, N. & Ohkuma, S. Autophagosome-lysosome fusion depends on the pH in acidic compartments in CHO cells. Autophagy 3, 154–157 (2007). · DOI
  43. Mangieri, L. R. et al. ATP6V0C knockdown in neuroblastoma cells alters autophagy-lysosome pathway function and metabolism of proteins that accumulate in neurodegenerative disease. PLoS ONE 9, e93257 (2014). · DOI
  44. Sobota, J. A., Back, N., Eipper, B. A. & Mains, R. E. Inhibitors of the V0 subunit of the vacuolar H+-ATPase prevent segregation of lysosomal- and secretory-pathway proteins. J. Cell Sci. 122, 3542–3553 (2009). · DOI
  45. Teplova, V. V., Tonshin, A. A., Grigoriev, P. A., Saris, N. E. & Salkinoja-Salonen, M. S. Bafilomycin A1 is a potassium ionophore that impairs mitochondrial functions. J. Bioenerg. Biomembr. 39, 321–329 (2007). · DOI
  46. Shacka, J. J. et al. Bafilomycin A1 inhibits chloroquine-induced death of cerebellar granule neurons. Mol. Pharmacol. 69, 1125–1136 (2006). · DOI
  47. Lopez, J. J., Camello-Almaraz, C., Pariente, J. A., Salido, G. M. & Rosado, J. A. Ca2+ accumulation into acidic organelles mediated by Ca2+- and vacuolar H+-ATPases in human platelets. Biochem. J. 390, 243–252 (2005). · DOI
  48. Goldman, F. D. et al. Hydroxychloroquine inhibits calcium signals in T cells: a new mechanism to explain its immunomodulatory properties. Blood 95, 3460–3466 (2000). · DOI
  49. Minelli, A., Lyons, S., Nolte, C., Verkhratsky, A. & Kettenmann, H. Ammonium triggers calcium elevation in cultured mouse microglial cells by initiating Ca(2+) release from thapsigargin-sensitive intracellular stores. Pflugers Arch. 439, 370–377 (2000).
  50. Parkinson, K. et al. Calcium-dependent regulation of Rab activation and vesicle fusion by an intracellular P2X ion channel. Nat. Cell. Biol. 16, 87–98 (2014). · DOI
  51. Jena, B. P. Role of SNAREs in membrane fusion. Adv. Exp. Med. Biol. 713, 13–32 (2011). · DOI
  52. Hay, J. C. Calcium: a fundamental regulator of intracellular membrane fusion? EMBO Rep. 8, 236–240 (2007). · DOI
  53. Kilpatrick, B. S., Eden, E. R., Schapira, A. H., Futter, C. E. & Patel, S. Direct mobilisation of lysosomal Ca2+ triggers complex Ca2+ signals. J. Cell Sci. 126, 60–66 (2013). · DOI
  54. Morgan, A. J. et al. Bidirectional Ca(2)(+) signaling occurs between the endoplasmic reticulum and acidic organelles. J. Cell Biol. 200, 789–805 (2013). · DOI
  55. Lopez-Sanjurjo, C. I., Tovey, S. C., Prole, D. L. & Taylor, C. W. Lysosomes shape Ins(1,4,5)P3-evoked Ca2+ signals by selectively sequestering Ca2+ released from the endoplasmic reticulum. J. Cell Sci. 126, 289–300 (2013). · DOI
  56. Wong, C. O., Li, R., Montell, C. & Venkatachalam, K. Drosophila TRPML is required for TORC1 activation. Curr. Biol. 22, 1616–1621 (2012). · DOI
  57. Park, H. W. et al. Pharmacological correction of obesity-induced autophagy arrest using calcium channel blockers. Nat. Commun. 5, 4834 (2014). · DOI
  58. Decuypere, J. P., Bultynck, G. & Parys, J. B. A dual role for Ca(2+) in autophagy regulation. Cell Calcium 50, 242–250 (2011). · DOI
  59. Arsham, A. M. & Neufeld, T. P. A genetic screen in Drosophila reveals novel cytoprotective functions of the autophagy-lysosome pathway. PLoS ONE 4, e6068 (2009). · DOI
  60. Zhang, J. et al. Thirty-one flavors of Drosophila rab proteins. Genetics 176, 1307–1322 (2007). · DOI
  61. Buechling, T. et al. Wnt/Frizzled signaling requires dPRR, the Drosophila homolog of the prorenin receptor. Curr. Biol. 20, 1263–1268 (2010). · DOI
  62. Pulipparacharuvil, S. et al. Drosophila Vps16A is required for trafficking to lysosomes and biogenesis of pigment granules. J. Cell. Sci. 118, 3663–3673 (2005). · DOI
  63. Sun, X. R. et al. Fast GCaMPs for improved tracking of neuronal activity. Nat. Commun. 4, 2170 (2013). · DOI
  64. Hennig, K. M., Colombani, J. & Neufeld, T. P. TOR coordinates bulk and targeted endocytosis in the Drosophila melanogaster fat body to regulate cell growth. J. Cell Biol. 173, 963–974 (2006). · DOI

Cited by

Sorted by citations

See all 424

Metadata

Authors
Mauvezin, C., Nagy, P., Juhász, G., & Neufeld, T. P.
ORCID
Mauvezin, Caroline
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2015-05-11
Volume
6
Issue
1
ISSN
2041-1723
DOI
10.1038/ncomms8007
PMC
PMC4428688
Cited by
424
References
6
Language
en
Licence

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review