Vacuolar ATPase in phagosome-lysosome fusion

Sandra Kissing, Christina Hermsen, Urska Repnik, Cecilie Kåsi Nesset, Kristine Von Bargen, Gareth Griffiths, Atsuhiro Ichihara, Beth S. Lee, Michael Schwake, Jef De Brabander, Albert Haas, Paul Saftig

Research output: Contribution to journalArticle

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Abstract

The vacuolar H<sup>+</sup>-ATPase (v-ATPase) complex is instrumental in establishing and maintaining acidification of some cellular compartments, thereby ensuring their functionality. Recently it has been proposed that the transmembrane V<inf>0</inf> sector of v-ATPase and its a-subunits promote membrane fusion in the endocytic and exocytic pathways independent of their acidification functions. Here, we tested if such a proton-pumping independent role of v-ATPase also applies to phagosome-lysosome fusion. Surprisingly, endo(lyso)somes in mouse embryonic fibroblasts lacking the V<inf>0</inf> a3 subunit of the v-ATPase acidified normally, and endosome and lysosome marker proteins were recruited to phagosomes with similar kinetics in the presence or absence of the a3 subunit. Further experiments used macrophages with a knockdown of v-ATPase accessory protein 2 (ATP6AP2) expression, resulting in a strongly reduced level of the V<inf>0</inf> sector of the v-ATPase. However, acidification appeared undisturbed, and fusion between latex bead-containing phagosomes and lysosomes, as analyzed by electron microscopy, was even slightly enhanced, as was killing of non-pathogenic bacteria by V<inf>0</inf> mutant macrophages. Pharmacologically neutralized lysosome pH did not affect maturation of phagosomes in mouse embryonic cells or macrophages. Finally, locking the two large parts of the v-ATPase complex together by the drug saliphenylhalamide A did not inhibit in vitro and in cellulo fusion of phagosomes with lysosomes. Hence, our data do not suggest a fusion-promoting role of the v-ATPase in the formation of phagolysosomes.

Original languageEnglish (US)
Pages (from-to)14166-14180
Number of pages15
JournalJournal of Biological Chemistry
Volume290
Issue number22
DOIs
StatePublished - May 29 2015

Fingerprint

Vacuolar Proton-Translocating ATPases
Phagosomes
Lysosomes
Fusion reactions
Acidification
Macrophages
Proton-Translocating ATPases
Membrane Fusion
Endosomes
Latex
Accessories
Fibroblasts
Microspheres
Electron microscopy
Protons
Electron Microscopy
Bacteria
Proteins
Membranes
Kinetics

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Kissing, S., Hermsen, C., Repnik, U., Nesset, C. K., Von Bargen, K., Griffiths, G., ... Saftig, P. (2015). Vacuolar ATPase in phagosome-lysosome fusion. Journal of Biological Chemistry, 290(22), 14166-14180. https://doi.org/10.1074/jbc.M114.628891

Vacuolar ATPase in phagosome-lysosome fusion. / Kissing, Sandra; Hermsen, Christina; Repnik, Urska; Nesset, Cecilie Kåsi; Von Bargen, Kristine; Griffiths, Gareth; Ichihara, Atsuhiro; Lee, Beth S.; Schwake, Michael; De Brabander, Jef; Haas, Albert; Saftig, Paul.

In: Journal of Biological Chemistry, Vol. 290, No. 22, 29.05.2015, p. 14166-14180.

Research output: Contribution to journalArticle

Kissing, S, Hermsen, C, Repnik, U, Nesset, CK, Von Bargen, K, Griffiths, G, Ichihara, A, Lee, BS, Schwake, M, De Brabander, J, Haas, A & Saftig, P 2015, 'Vacuolar ATPase in phagosome-lysosome fusion', Journal of Biological Chemistry, vol. 290, no. 22, pp. 14166-14180. https://doi.org/10.1074/jbc.M114.628891
Kissing S, Hermsen C, Repnik U, Nesset CK, Von Bargen K, Griffiths G et al. Vacuolar ATPase in phagosome-lysosome fusion. Journal of Biological Chemistry. 2015 May 29;290(22):14166-14180. https://doi.org/10.1074/jbc.M114.628891
Kissing, Sandra ; Hermsen, Christina ; Repnik, Urska ; Nesset, Cecilie Kåsi ; Von Bargen, Kristine ; Griffiths, Gareth ; Ichihara, Atsuhiro ; Lee, Beth S. ; Schwake, Michael ; De Brabander, Jef ; Haas, Albert ; Saftig, Paul. / Vacuolar ATPase in phagosome-lysosome fusion. In: Journal of Biological Chemistry. 2015 ; Vol. 290, No. 22. pp. 14166-14180.
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