Phosphatidylserine dictates the assembly and dynamics of caveolae in the plasma membrane

Hirama, Takashi, Das, Raibatak, Yang, Yanbo, Ferguson, Charles, Won, Amy, Yip, Christopher M., Kay, Jason G., Grinstein, Sergio, Parton, Robert G. and Fairn, Gregory D. (2017) Phosphatidylserine dictates the assembly and dynamics of caveolae in the plasma membrane. Journal of Biological Chemistry, 292 34: 14292-14307. doi:10.1074/jbc.M117.791400

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Author Hirama, Takashi
Das, Raibatak
Yang, Yanbo
Ferguson, Charles
Won, Amy
Yip, Christopher M.
Kay, Jason G.
Grinstein, Sergio
Parton, Robert G.
Fairn, Gregory D.
Title Phosphatidylserine dictates the assembly and dynamics of caveolae in the plasma membrane
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 1083-351X
0021-9258
Publication date 2017-08-25
Year available 2017
Sub-type Critical review of research, literature review, critical commentary
DOI 10.1074/jbc.M117.791400
Open Access Status File (Publisher version)
Volume 292
Issue 34
Start page 14292
End page 14307
Total pages 16
Place of publication Rockville, MD United States
Publisher American Society for Biochemistry and Molecular Biology
Language eng
Abstract Caveolae are bulb-shaped nanodomains of the plasma membrane that are enriched in cholesterol and sphingolipids. They have many physiological functions, including endocytic transport, mechanosensing, and regulation of membrane and lipid transport. Caveola formation relies on integral membrane proteins termed caveolins (Cavs) and the cavin family of peripheral proteins. Both protein families bind anionic phospholipids, but the precise roles of these lipids are unknown. Here, we studied the effects of phosphatidylserine (PtdSer), phosphatidylinositol 4-phosphate (PtdIns4P), and phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) on caveolar formation and dynamics. Using live-cell, single-particle tracking of GFP-labeled Cav1 and ultrastructural analyses, we compared the effect of PtdSer disruption or phosphoinositide depletion with caveola disassembly caused by cavin1 loss. We found that PtdSer plays a crucial role in both caveola formation and stability. Sequestration or depletion of PtdSer decreased the number of detectable Cav1-GFP puncta and the number of caveolae visualized by electron microscopy. Under PtdSer-limiting conditions, the co-localization of Cav1 and cavin1 was diminished, and cavin1 degradation was increased. Using rapamycin-recruitable phosphatases, we also found that the acute depletion of PtdIns4P and PtdIns(4,5)P2 has minimal impact on caveola assembly but results in decreased lateral confinement. Finally, we show in a model of phospholipid scrambling, a feature of apoptotic cells, that caveola stability is acutely affected by the scrambling. We conclude that the predominant plasmalemmal anionic lipid PtdSer is essential for proper Cav clustering, caveola formation, and caveola dynamics and that membrane scrambling can perturb caveolar stability.
Formatted abstract
Caveolae are bulb-shaped nanodomains of the plasma membrane that are enriched in cholesterol and sphingolipids. They have many physiological functions, including endocytic transport, mechanosensing, and regulation of membrane and lipid transport. Caveola formation relies on integral membrane proteins termed caveolins (Cavs) and the cavin family of peripheral proteins. Both protein families bind anionic phospholipids, but the precise roles of these lipids are unknown. Here, we studied the effects of phosphatidylserine (PtdSer), phosphatidylinositol 4-phosphate (PtdIns4P), and phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) on caveolar formation and dynamics. Using live-cell, single-particle tracking of GFP-labeled Cav1 and ultrastructural analyses, we compared the effect of PtdSer disruption or phosphoinositide depletion with caveola disassembly caused by cavin1 loss. We found that PtdSer plays a crucial role in both caveola formation and stability. Sequestration or depletion of PtdSer decreased the number of detectable Cav1-GFP puncta and the number of caveolae visualized by electron microscopy. Under PtdSer-limiting conditions, the co-localization of Cav1 and cavin1 was diminished, and cavin1 degradation was increased. Using rapamycin-recruitable phosphatases, we also found that the acute depletion of PtdIns4P and PtdIns(4,5)P2 has minimal impact on caveola assembly but results in decreased lateral confinement. Finally, we show in a model of phospholipid scrambling, a feature of apoptotic cells, that caveola stability is acutely affected by the scrambling. We conclude that the predominant plasmalemmal anionic lipid PtdSer is essential for proper Cav clustering, caveola formation, and caveola dynamics and that membrane scrambling can perturb caveolar stability.
Keyword Caveolae
Microscopic imaging
Phosphatidylinositol signaling
Phosphatidylserine
Single-particle analysis
Phosphoinositides
TIRF microscopy
Single-particle analysis
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID 1
Institutional Status UQ
Additional Notes This research was originally published in Journal of Biological Chemistry. Hirama, Takashi, Das, Raibatak, Yang, Yanbo, Ferguson, Charles, Won, Amy, Yip, Christopher M., Kay, Jason G., Grinstein, Sergio, Parton, Robert G. and Fairn, Gregory D. Phosphatidylserine dictates the assembly and dynamics of caveolae in the plasma membrane. Journal of Biological Chemistry. 2017; 292:14292-14307. © the American Society for Biochemistry and Molecular Biology.

Document type: Journal Article
Sub-type: Critical review of research, literature review, critical commentary
Collections: HERDC Pre-Audit
Institute for Molecular Bioscience - Publications
Centre for Microscopy and Microanalysis Publications
 
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