Lipid kinases VPS34 and PIKfyve coordinate a phosphoinositide cascade to regulate Retriever-mediated recycling on endosomes
- University of Michigan-Ann Arbor, United States
- INSERM U1048 I2MC, France
- The University of Texas Southwestern Medical Center, United States
- Mayo Clinic, United States
Abstract
Cell-surface receptors control how cells respond to their environment. Many cell-surface receptors recycle from endosomes to the plasma membrane via a recently discovered pathway, which includes sorting-nexin SNX17, Retriever, WASH and CCC complexes. Here, using mammalian cells, we discover that PIKfyve and its upstream PI3-kinase VPS34 positively regulate this pathway. VPS34 produces PI3P, which is the substrate for PIKfyve to generate PI3,5P2. We show that PIKfyve controls recycling of cargoes including integrins, receptors that control cell migration. Furthermore, endogenous PIKfyve colocalizes with SNX17, Retriever, WASH and CCC complexes on endosomes. Importantly, PIKfyve inhibition results displacement of Retriever and CCC from endosomes. In addition, we show that recruitment of SNX17 is an early step and requires VPS34. These discoveries suggest that VPS34 and PIKfyve coordinate an ordered pathway to regulate recycling from endosomes and suggest how PIKfyve functions in cell migration.
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All data generated or analyzed during this study are included in the manuscript and supporting files.
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Funding
National Institute of Neurological Disorders and Stroke (Research Project Grant R01 NS064015)
- Lois S Weisman
National Institute of Neurological Disorders and Stroke (Research Project Grant R01-NS099340)
- Lois S Weisman
National Institute of Diabetes and Digestive and Kidney Diseases (Research Project Grant R01-DK107733)
- Ezra Burstein
- Daniel D Billadeau
American Heart Association (Postdoctoral Fellowship 14POST20480137)
- Sai Srinivas Panapakkam Giridharan
American Heart Association (Postdoctoral Fellowship 19POST34450253)
- Guangming Luo
University of Michigan Protein Folding Diseases Fast Forward Initiative (Pilot grant)
- Lois S Weisman
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (#A3114-01) of the University of Michigan. The protocol was approved by the Committee on the Ethics of Animal Experiments of the University of Michigan (Approval # PRO00010100).
Copyright
© 2022, Giridharan et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Lipid kinases VPS34 and PIKfyve coordinate a phosphoinositide cascade to regulate Retriever-mediated recycling on endosomes
eLife 11:e69709.
https://doi.org/10.7554/eLife.69709
