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Acetate and hypertonic stress stimulate organelle membrane fission using distinct phosphatidylinositol signals

Dipti Patel, View ORCID ProfileChristopher Leonard Brett
doi: https://doi.org/10.1101/398685
Dipti Patel
1Concordia University, Department of Biology, 7141 Sherbrooke St. W., SP-501.15, Montreal, QC, H4B 1R6, Canada
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Christopher Leonard Brett
1Concordia University, Department of Biology, 7141 Sherbrooke St. W., SP-501.15, Montreal, QC, H4B 1R6, Canada
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  • ORCID record for Christopher Leonard Brett
  • For correspondence: christopher.brett@concordia.ca
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ABSTRACT

Organelle morphology reflects an equilibrium between membrane fusion and fission that determines size, shape and copy number. By studying the yeast vacuole as a model, the conserved molecular mechanisms responsible for organelle fusion have been revealed. However, a detailed understanding of vacuole fission and how these opposing processes respond to the cell cycle, osmoregulation or metabolism to change morphology remain elusive. Thus, herein we describe a new fluorometric assay to measure vacuole fission in vitro. For proof-of-concept, we use this assay to confirm that acetate, a key intermediary metabolite, triggers vacuole fission in vitro and show that it also blocks homotypic vacuole fusion. The basis of this effect is distinct from hypertonic stress, a known trigger of fission and inhibitor of fusion that inactivates the Rab-GTPase Ypt7: Treatment with the phosphatidylinositol-kinase inhibitor wortmannin or the catalytic domain of the Rab-GAP (GTPase Activating Protein) Gyp1 reveal that fission can be triggered by Ypt7 inactivation alone in absence of hypertonic stress, placing it upstream of PI-3,5-P2 synthesis and osmosis required for membrane scission. Whereas acetate seems to block PI-4-kinase to possibly increase the pool of PI on vacuole membranes needed to synthesize sufficient PI-3,5-P2 for fission. Thus, we speculate that both PI-4-P and PI-3-P arms of PI-P signaling drive changes in membrane fission and fusion responsible altering vacuole morphology in response to cellular metabolism or osmoregulation.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted August 23, 2018.
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Acetate and hypertonic stress stimulate organelle membrane fission using distinct phosphatidylinositol signals
Dipti Patel, Christopher Leonard Brett
bioRxiv 398685; doi: https://doi.org/10.1101/398685
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Acetate and hypertonic stress stimulate organelle membrane fission using distinct phosphatidylinositol signals
Dipti Patel, Christopher Leonard Brett
bioRxiv 398685; doi: https://doi.org/10.1101/398685

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