Branched actin networks are organized for asymmetric force production during clathrin-mediated endocytosis in mammalian cells

Jin, Meiyan; Shirazinejad, Cyna; Wang, Bowen; Yan, Aiming; Schöneberg, Johannes; Upadhyayula, Srigokul; Xu, Ke; Drubin, David G.

doi:10.1038/s41467-022-31207-5

Cited by 22 publications

(17 citation statements)

References 54 publications

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“…Increased membrane tension reduces the initiation density ( Ferguson et al, 2017 ), and slows down the growth and dissolution rates of endocytic clathrin-coated structures ( Ferguson et al, 2016 ). Moreover, internalization of CPs from the plasma membrane becomes dependent on the curvature generating adaptor proteins ( Joseph et al, 2020 ; Willy et al, 2021a ) and the forces provided by actin polymerization under increased tension ( Boulant et al, 2011 ; Jin et al, 2022 ; Kaplan et al, 2022 ).…”

Section: Introductionmentioning

confidence: 99%

Endocytosis at extremes: Formation and internalization of giant clathrin-coated pits under elevated membrane tension

Akatay

Djakbarova

et al. 2022

Front. Mol. Biosci.

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Internalization of clathrin-coated vesicles from the plasma membrane constitutes the major endocytic route for receptors and their ligands. Dynamic and structural properties of endocytic clathrin coats are regulated by the mechanical properties of the plasma membrane. Here, we used conventional fluorescence imaging and multiple modes of structured illumination microscopy (SIM) to image formation of endocytic clathrin coats within live cells and tissues of developing fruit fly embryos. High resolution in both spatial and temporal domains allowed us to detect and characterize distinct classes of clathrin-coated structures. Aside from the clathrin pits and plaques detected in distinct embryonic tissues, we report, for the first time, formation of giant coated pits (GCPs) that can be up to two orders of magnitude larger than the canonical pits. In cultured cells, we show that GCP formation is induced by increased membrane tension. GCPs take longer to grow but their mechanism of curvature generation is the same as the canonical pits. We also demonstrate that GCPs split into smaller fragments during internalization. Considering the supporting roles played by actin filament dynamics under mechanically stringent conditions that slow down completion of clathrin coats, we suggest that local changes in the coat curvature driven by actin machinery can drive splitting and internalization of GCPs.

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Section: Introductionmentioning

confidence: 99%

Endocytosis at extremes: Formation and internalization of giant clathrin-coated pits under elevated membrane tension

Akatay

Djakbarova

et al. 2022

Front. Mol. Biosci.

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“…Our results suggest that CCSs on tracks experience a transient period of frustration, as reflected by their longer lifetime, but eventually manage to internalize portions of tracks. Actin has been proposed to be recruited at frustrated CCSs that experience difficulties to bud because of mechanical constraints (Baschieri et al, 2020; Boulant et al, 2011; Jin et al, 2022; Kaksonen et al, 2006; Perrais & Merrifield, 2005). Indeed, we observed that inhibiting actin dynamics with Cytochalasin D resulted in an increased accumulation of stable CCSs along tracks (Supp.…”

Section: Resultsmentioning

confidence: 99%

Fibroblasts generate topographical cues that steer cancer cell migration

Baschieri

Illand

Barbazán

et al. 2022

Preprint

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Fibroblasts play a fundamental role in tumor development. Among other functions, they regulate cancer cells migration through rearranging the extracellular matrix, secreting soluble factors and establishing direct physical contacts with cancer cells. Here, we report that migrating fibroblasts deposit on the substrate a network of tubular structures that serves as guidance cue for cancer cell migration. Such membranous tubular network, hereafter called tracks, is stably anchored to the substrate in a β5 integrin-dependent manner. We found that cancer cells specifically adhere to tracks by using clathrin-coated structures that pinch and engulf tracks. Tracks represent thus a spatial memory of fibroblast migration paths that is read and erased by cancer cells directionally migrating along them. We propose that fibroblast tracks represent a topography-based intercellular communication system capable of steering cancer cells migration.

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“…As a first step, we can check simulation times against the physical timescales of the system. Fluctuations of the membrane, with the wave vector q, have a hydrodynamic dissipation timescale of 4η/κq 3 , where η is the viscosity of the solvent and κ ≈ 20 kT is the bending rigidity of the membrane. This gives the hydrodynamic dissipation timescale to be in the 0.01 ms range.…”

Section: Resultsmentioning

confidence: 99%

Formation of membrane invaginations by curvature-inducing peripheral proteins: free energy profiles, kinetics, and membrane-mediated effects

Sadeghi

2022

Preprint

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Curvature-inducing peripheral proteins have been observed to spontaneously remodel bilayer membranes, resulting in membrane invaginations and formation of membrane tubules. In case of proteins such as cholera and Shiga toxins that bend the membrane with locally isotropic curvatures, the resulting membrane-mediated interactions are rather small. Thus, the process in which these proteins form dense clusters on the membrane and collectively induce invaginations is extremely slow, progressing over several minutes. This makes it virtually impossible to directly simulate the pathway leading to membrane tubulation even with highly coarse-grained models. Here, we present a steered molecular dynamics protocol through which the peripheral proteins are forced to gather on a membrane patch and form a tubular invagination. Using thermodynamic integration, we obtain the free energy profile of this process and discuss its different stages. We show how protein stiffness, which also determines local membrane curvatures, affects the free energy landscape and the organization of proteins in the invaginated region. Furthermore, we estimate the kinetics of the described pathway modeled as a Markovian stochastic process, and compare the implied timescales with their experimental counterparts.

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Branched actin networks are organized for asymmetric force production during clathrin-mediated endocytosis in mammalian cells

Cited by 22 publications

References 54 publications

Endocytosis at extremes: Formation and internalization of giant clathrin-coated pits under elevated membrane tension

Endocytosis at extremes: Formation and internalization of giant clathrin-coated pits under elevated membrane tension

Fibroblasts generate topographical cues that steer cancer cell migration

Formation of membrane invaginations by curvature-inducing peripheral proteins: free energy profiles, kinetics, and membrane-mediated effects

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