Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae

Gambin, Yann; Ariotti, Nicholas; McMahon, Kerrie‐Ann; Bastiani, Michele; Sierecki, Emma; Kovtun, Oleksiy; Polinkovsky, Mark; Magenau, Astrid; Jung, WooRam; Okano, Satomi; Zhou, Yong; Leneva, Natalya; Mureev, Sergey; Johnston, Wayne A.; Gaus, Katharina; Hancock, John F.; Collins, Brett M.; Alexandrov, Kirill; Parton, Robert G.

doi:10.7554/elife.01434

Cited by 126 publications

(182 citation statements)

References 37 publications

(38 reference statements)

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“…1A). Using pulldown experiments we found that cavin1 self-associated and bound to cavin2 and cavin3, as previously suggested (Gambin et al, 2014), but that cavin2 and cavin3 did not interact ( Fig. 1B; supplementary material Fig.…”

Section: Resultssupporting

confidence: 48%

Cavin3 interacts with cavin1 and caveolin1 to increase surface dynamics of caveolae

Mohan

Morén

Larsson

et al. 2015

Journal of Cell Science

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Caveolae are invaginations of the cell surface thought to regulate membrane tension, signalling, adhesion and lipid homeostasis owing to their dynamic behaviour ranging from stable surface association to dynamic rounds of fission and fusion with the plasma membrane. The caveolae coat is generated by oligomerisation of the membrane protein caveolin and the family of cavin proteins. Here, we show that cavin3 (also known as PRKCDBP) is targeted to caveolae by cavin1 (also known as PTRF) where it interacts with the scaffolding domain of caveolin1 and promote caveolae dynamics. We found that the N-terminal region of cavin3 binds a trimer of the cavin1 N-terminus in competition with a homologous cavin2 (also known as SDPR) region, showing that the cavins form distinct subcomplexes through their N-terminal regions. Our data shows that cavin3 is enriched at deeply invaginated caveolae and that loss of cavin3 in cells results in an increase of stable caveolae and a decrease of caveolae that are only present at the membrane for a short time. We propose that cavin3 is recruited to the caveolae coat by cavin1 to interact with caveolin1 and regulate the duration time of caveolae at the plasma membrane.

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Section: Resultssupporting

confidence: 48%

Cavin3 interacts with cavin1 and caveolin1 to increase surface dynamics of caveolae

Mohan

Morén

Larsson

et al. 2015

Journal of Cell Science

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“…The cavins also have the ability to bind to PtdSer (16,17,20). Furthermore, a recent study demonstrated that there are 50 cavin1 molecules per caveola (15,21). Together these observations suggest that PtdSer may play a critical role in the formation and stabilization of caveolae.…”

Section: Edited By Dennis R Voelkermentioning

confidence: 66%

“…In the cellular context at first glance, the strength of the interaction between three cationic amino acids and the anionic phospholipid would be rather modest. However, as caveolae have been estimated to contain Ϸ144 Cav1 molecules (9,15), the number of basic residues per caveola is amplified to Ϸ432. Thus, the potential strength of the electrostatic interaction is considerable.…”

Section: Edited By Dennis R Voelkermentioning

confidence: 99%

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

Hirama

Das

Yang

et al. 2017

Journal of Biological Chemistry

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Edited by Dennis R. VoelkerCaveolae 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)P 2 ) 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)P 2 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.

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“…24 This network of low affinity interactions presumably allows disassembly of caveolae in response to increased membrane tension as cavins are released from caveolae as they flatten. In addition, with an estimated 140-150 caveolin molecules 25 and 50 cavin1 molecules 26 in a single caveolae the lipid domain generated within caveolae may be quite distinct to that of the bulk plasma membrane. The biophysical properties of the caveolar domain generated by the combination of caveolin insertion into the membrane, the cytoplasmic coat of cavin proteins, and their distinct lipid composition might be very different to that of the bulk membrane but at present the precise quantitative parameters are unknown.…”

mentioning

confidence: 99%

Mechanoprotection by skeletal muscle caveolae

Hall

Parton

2016

BioArchitecture

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ABSTRACT. Caveolae, small bulb-like pits, are the most abundant surface feature of many vertebrate cell types. The relationship of the structure of caveolae to their function has been a subject of considerable scientific interest in view of the association of caveolar dysfunction with human disease. In a recent study Lo et al.1 investigated the organization and function of caveolae in skeletal muscle. Using quantitative 3D electron microscopy caveolae were shown to be predominantly organized into multilobed structures which provide a large reservoir of surface-connected membrane underlying the sarcolemma. These structures were preferentially disassembled in response to changes in membrane tension. Perturbation or loss of caveolae in mouse and zebrafish models suggested that caveolae can protect the muscle sarcolemma against damage in response to excessive membrane activity. Flattening of caveolae to release membrane into the bulk plasma membrane in response to increased membrane tension can allow cell shape changes and prevent membrane rupture. In addition, disassembly of caveolae can have widespread effects on lipid-based plasma membrane organization. These findings suggest that the ability of the caveolar membrane system to respond to mechanical forces is a crucial evolutionarily-conserved process which is compromised in disease conditions associated with mutations in key caveolar components.KEYWORDS. caveolae, mechanoprotection, skeletal muscle, T-tubule ABBREVIATIONS. PS, phosphatidylserine; PI(4,5)P 2 , phosphatidylinositol 4,5 bisphosphateThe sarcolemma of the skeletal muscle fiber is a highly specialized structure, which must cope with the physical demands of muscle contraction, respond to extracellular signals, and transmit action potentials from the surface of the muscle to the interior. These functions are facilitated by differentiation of the muscle surface into distinct macrodomains; the sarcolemma, with associated microdomains such as caveolae and clathrin coated pits, and a second extensive domain, the T-tubule system, itself possessing distinct microdomains termed triads specialized for electrical coupling to the sarcoplasmic muscle to allow synchronized calcium release. Understanding how these membrane systems are generated, maintained, and their role in muscle function is crucial as numerous human disease conditions are associated with

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Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae

Cited by 126 publications

References 37 publications

Cavin3 interacts with cavin1 and caveolin1 to increase surface dynamics of caveolae

Cavin3 interacts with cavin1 and caveolin1 to increase surface dynamics of caveolae

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

Mechanoprotection by skeletal muscle caveolae

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