Cell-free formation and interactome analysis of caveolae

Jung, WooRam; Sierecki, Emma; Bastiani, Michele; O’Carroll, Ailís; Alexandrov, Kirill; Rae, James; Johnston, Wayne A.; Hunter, Dominic J. B.; Ferguson, Charles; Gambin, Yann; Ariotti, Nicholas; Parton, Robert G.

doi:10.1083/jcb.201707004

Cited by 52 publications

(62 citation statements)

References 95 publications

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“…A, Cav-a-tGFP localizes at the peri-nucleus in Ciona embryos at 14 hpf. 17,18 In the urochordate animal Ciona, our results using a cell-free system, as well as in vivo experimental results, revealed that Cav-a was recruited to the plasma membrane, and generated membrane invaginations that were morphologically similar to vertebrate caveolae, but with a wider "neck". C, At 21 hpf, Cav-a-tGFP specially enriches at the apical domains.…”

Section: Ascidian Cav-a Forms Membrane Curvature In Both In Vitro Amentioning

confidence: 70%

“…18 The results showed that Ciona Cav-a but not Cav-b induced membrane curvature similar to that induced by human Cav1 (red circle in Figure 3A). 18 The results showed that Ciona Cav-a but not Cav-b induced membrane curvature similar to that induced by human Cav1 (red circle in Figure 3A).…”

Section: Cav-a Forms Caveola-like Structures By Inducing Membrane Cmentioning

confidence: 78%

“…18 For Immuno-gold labeling, the Glow-discharged carbon coated grid was placed on a sample droplet for 20 minutes, followed by blocking with 1% casein in PBS 3 times for 5 minutes. 18 For Immuno-gold labeling, the Glow-discharged carbon coated grid was placed on a sample droplet for 20 minutes, followed by blocking with 1% casein in PBS 3 times for 5 minutes.…”

Section: Cell-free Expression Experimentsmentioning

confidence: 99%

“…11 It is worth noting that cavins are absent in the invertebrate genome. 14,15 Only Cav1 and Cav3 can drive caveola formation, [16][17][18] suggesting that Cav1 has a major function in nonmuscle cells. 13 Cav1 and Cav2 are coexpressed in many nonmuscle cells whereas Cav3 is muscle-specific and has a role in muscle development in vertebrates.…”

Section: Introductionmentioning

confidence: 99%

“…[20][21][22][23][24] Mutations of caveolin genes have been linked to a number of diseases in humans. 18 Until now, no visible caveolae structures have been observed in invertebrate tissues, 12 with the exception of the description of a caveola-like structure in C. elegans. 16 The expression of vertebrate caveolins in bacteria has been shown to form caveolae-like structures.…”

Section: Introductionmentioning

confidence: 99%

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Ascidian caveolin induces membrane curvature and protects tissue integrity and morphology during embryogenesis

Bhattachan

Rae

et al. 2019

The FASEB Journal

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Abbreviations: ANISEED, ascidian network for in situ expression and embryological data; APEX, soybean ascorbate peroxidase; BFA, brefeldin A; BHK, baby hamster kidney; Cav, caveolin; ECM, extracellular matrix; EF1α, elongation factor 1 alpha; EGFP, enhanced green fluorescent protein; FBS, fetal bovine serum; FRAP, fluorescence recovery after photobleaching AbstractCell morphology and tissue integrity are essential for embryogenesis. Caveolins are membrane proteins that induce the formation of surface pits called caveolae that serve as membrane reservoirs for cell and tissue protection during development. In vertebrates, caveolin 1 (Cav1) and caveolin 3 (Cav3) are required for caveola formation.

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Section: Ascidian Cav-a Forms Membrane Curvature In Both In Vitro Amentioning

confidence: 70%

Section: Cav-a Forms Caveola-like Structures By Inducing Membrane Cmentioning

confidence: 78%

Section: Cell-free Expression Experimentsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Ascidian caveolin induces membrane curvature and protects tissue integrity and morphology during embryogenesis

Bhattachan

Rae

et al. 2019

The FASEB Journal

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Adhesion‐dependent Caveolin‐1 Tyrosine‐14 phosphorylation is regulated by FAK in response to changing matrix stiffness

et al. 2021

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Integrin‐mediated adhesion regulates cellular responses to changes in the mechanical and biochemical properties of the extracellular matrix. Cell–matrix adhesion regulates caveolar endocytosis, dependent on caveolin 1 (Cav1) Tyr14 phosphorylation (pY14Cav1), to control anchorage‐dependent signaling. We find that cell–matrix adhesion regulates pY14Cav1 levels in mouse fibroblasts. Biochemical fractionation reveals endogenous pY14Cav1 to be present in caveolae and focal adhesions (FA). Adhesion does not affect caveolar pY14Cav1, supporting its regulation at FA, in which PF‐228‐mediated inhibition of focal adhesion kinase (FAK) disrupts. Cell adhesion on 2D polyacrylamide matrices of increasing stiffness stimulates Cav1 phosphorylation, which is comparable to the phosphorylation of FAK. Inhibition of FAK across varying stiffnesses shows it regulates pY14Cav1 more prominently at higher stiffness. Taken together, these studies reveal the presence of FAK–pY14Cav1 crosstalk at FA, which is regulated by cell–matrix adhesion.

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Single‐Molecule Counting Coupled to Rapid Amplification Enables Detection of α‐Synuclein Aggregates in Cerebrospinal Fluid of Parkinson's Disease Patients

et al. 2021

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α‐Synuclein aggregation is a hallmark of Parkinson's disease and a promising biomarker for early detection and assessment of disease progression. The prospect of a molecular test for Parkinson's disease is materializing with the recent developments of detection methods based on amplification of synuclein seeds (e.g. RT‐QuIC or PMCA). Here we adapted single‐molecule counting methods for the detection of α‐synuclein aggregates in cerebrospinal fluid (CSF), using a simple 3D printed microscope. Single‐molecule methods enable to probe the early events in the amplification process used in RT‐QuIC and a precise counting of ThT‐positive aggregates. Importantly, the use of single‐molecule counting also allows a refined characterization of the samples and fingerprinting of the protein aggregates present in CSF of patients. The fingerprinting of size and reactivity of individual aggregate shows a unique signature for each PD patients compared to controls and may provide new insights on synucleinopathies in the future.

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Cell-free formation and interactome analysis of caveolae

Cited by 52 publications

References 95 publications

Ascidian caveolin induces membrane curvature and protects tissue integrity and morphology during embryogenesis

Ascidian caveolin induces membrane curvature and protects tissue integrity and morphology during embryogenesis

Adhesion‐dependent Caveolin‐1 Tyrosine‐14 phosphorylation is regulated by FAK in response to changing matrix stiffness

Single‐Molecule Counting Coupled to Rapid Amplification Enables Detection of α‐Synuclein Aggregates in Cerebrospinal Fluid of Parkinson's Disease Patients

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