Copper blocks V-ATPase activity and SNARE complex formation to inhibit yeast vacuole fusion

Gregory E. Miner, Katherine D. Sullivan, Chi Zhang, Logan R. Hurst, Matthew L. Starr, David A. Rivera-Kohr, Brandon C. Jones, Annie Guo, Rutilio Fratti

Research output: Contribution to journalArticle

Abstract

The accumulation of copper in organisms can lead to altered functions of various pathways and become cytotoxic through the generation of reactive oxygen species. In yeast, cytotoxic metals such as Hg+, Cd2+ and Cu2+ are transported into the lumen of the vacuole through various pumps. Copper ions are initially transported into the cell by the copper transporter Ctr1 at the plasma membrane and sequestered by chaperones and other factors to prevent cellular damage by free cations. Excess copper ions can subsequently be transported into the vacuole lumen by an unknown mechanism. Transport across membranes requires the reduction of Cu2+ to Cu+. Labile copper ions can interact with membranes to alter fluidity, lateral phase separation and fusion. Here we found that CuCl2 potently inhibited vacuole fusion by blocking SNARE pairing. This was accompanied by the inhibition of V-ATPase H+ pumping. Deletion of the vacuolar reductase Fre6 had no effect on the inhibition of fusion by copper. This suggests that Cu2+ is responsible for the inhibition of vacuole fusion and V-ATPase function. This notion is supported by the differential effects of chelators. The Cu2+-specific chelator triethylenetetramine rescued fusion, whereas the Cu+-specific chelator bathocuproine disulfonate had no effect on the inhibited fusion.

Original languageEnglish (US)
Pages (from-to)841-850
Number of pages10
JournalTraffic
Volume20
Issue number11
DOIs
StatePublished - Nov 1 2019

Fingerprint

SNARE Proteins
Vacuoles
Yeast
Adenosine Triphosphatases
Copper
Fusion reactions
Yeasts
Chelating Agents
Ions
Trientine
Membranes
Proton-Translocating ATPases
Fluidity
Cell membranes
Phase separation
Cations
Reactive Oxygen Species
Oxidoreductases
Metals
Cell Membrane

Keywords

  • Fre6
  • Nyv1
  • SNARE
  • Sec17
  • V-ATPase
  • Vam3
  • Vam7
  • membrane fusion

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Miner, G. E., Sullivan, K. D., Zhang, C., Hurst, L. R., Starr, M. L., Rivera-Kohr, D. A., ... Fratti, R. (2019). Copper blocks V-ATPase activity and SNARE complex formation to inhibit yeast vacuole fusion. Traffic, 20(11), 841-850. https://doi.org/10.1111/tra.12683

Copper blocks V-ATPase activity and SNARE complex formation to inhibit yeast vacuole fusion. / Miner, Gregory E.; Sullivan, Katherine D.; Zhang, Chi; Hurst, Logan R.; Starr, Matthew L.; Rivera-Kohr, David A.; Jones, Brandon C.; Guo, Annie; Fratti, Rutilio.

In: Traffic, Vol. 20, No. 11, 01.11.2019, p. 841-850.

Research output: Contribution to journalArticle

Miner, GE, Sullivan, KD, Zhang, C, Hurst, LR, Starr, ML, Rivera-Kohr, DA, Jones, BC, Guo, A & Fratti, R 2019, 'Copper blocks V-ATPase activity and SNARE complex formation to inhibit yeast vacuole fusion', Traffic, vol. 20, no. 11, pp. 841-850. https://doi.org/10.1111/tra.12683
Miner GE, Sullivan KD, Zhang C, Hurst LR, Starr ML, Rivera-Kohr DA et al. Copper blocks V-ATPase activity and SNARE complex formation to inhibit yeast vacuole fusion. Traffic. 2019 Nov 1;20(11):841-850. https://doi.org/10.1111/tra.12683
Miner, Gregory E. ; Sullivan, Katherine D. ; Zhang, Chi ; Hurst, Logan R. ; Starr, Matthew L. ; Rivera-Kohr, David A. ; Jones, Brandon C. ; Guo, Annie ; Fratti, Rutilio. / Copper blocks V-ATPase activity and SNARE complex formation to inhibit yeast vacuole fusion. In: Traffic. 2019 ; Vol. 20, No. 11. pp. 841-850.
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