Endocytosis frustration potentiates compression-induced receptor signaling

Abstract

Cells experience mechanical stresses in different physiological and pathological settings.Clathrin-coated structures (CCSs) are sensitive to such perturbations in a way that often results in a mechanical impairment of their capacity to bud, ultimately impairing endocytosis.Compressive stress is a particular mechanical perturbation that leads to increased membrane tension and promotes proliferative signals. Here, we report that compression leads to CCSs frustration and that CCSs are required to potentiate receptor-mediated signaling in these conditions. We first confirmed that pressure stalls CCSs dynamics and showed that it also slows down the dynamic exchange of CCSs building blocks. As previously reported, compressioninduced paracrine activation of the epidermal growth factor receptor (EGFR) was the primary cause of ERK activation in these conditions. We observed that the EGFR was efficiently recruited at CCSs upon compression and that CCSs were required for full ERK activation. In addition, we demonstrated that compression-induced frustrated CCSs could also serve as signaling platforms for the hepatocyte growth factor receptor (HGFR), provided HGF was present in the medium. We thus propose that, besides the particular case of EGFR paracrine activation, CCS frustration resulting from mechanical perturbations can potentiate signaling through different receptors with potential important consequences on cell adaptation to its environment.

Plasma membrane nanodeformations promote actin polymerisation through CIP4/CDC42 recruitment and regulate type II IFN signaling

“…This could increase the probability for IFNγ-R of being trapped, preventing its signaling. Additionally, it is tempting to establish a direct conceptual analogy between the FEME-like structures described in our study and frustrated clathrin-mediated endocytosis described by other groups, where clathrin plaques were shown to act as platforms regulating receptor signaling (in particular for epidermal growth factor receptor) [22–24]. Yet, additional investigations are necessary to characterise IFNγ-R dynamics within these topography-induced actin nanodomains.…”

“…Furthermore, using correlative FluidFM/fast-scanning confocal microscopy, we demonstrated that these actin structures can form both at basal and apical plasma membranes, in a very dynamic manner. In addition, these membrane nanodomains are reminiscent of early-stage FEME-like endocytic pits, opening up the exciting possibility of the existence of frustrated clathrin-independent endocytosis, as has been shown with the frustrated clathrin-mediated endocytosis previously described [22][23][24]. Furthermore, these actin-enriched nanodomains harbour Interferon γ receptors (IFNγ-R) with an impaired signaling.…”

The Hippo pathway drives the cellular response to hydrostatic pressure

“…In CDE, a range of adaptor proteins, including AP2, actin filaments and the ANTH domain containing protein AP180, are recruited to the plasma membrane and cause formation of clathrin coats (Pearse, 1976) in a precise and temporal manner (Ford et al , 2001; Taylor et al , 2011; Yoshida et al , 2018; Akamatsu et al , 2020), where after the coated vesicles with internalized cargo are pinched off by the recruitment of the large GTPase dynamin (Ford et al , 2001; Taylor et al , 2011; Yoshida et al , 2018; Akamatsu et al , 2020). Importantly, CDE is mechanosensitive (Boulant et al , 2011; Saleem et al , 2015; Ferguson et al , 2017; Malinverno et al , 2017; Akamatsu et al , 2020; Baschieri et al , 2020) and in non‐specialized cells account for upwards of 95% internalization of the plasma membrane (Bitsikas et al , 2014). CDE therefore represents the major route for internalization of the plasma membrane and the process is an attractive pathway that could confer the change in cell surface plasma membrane necessary for changing the cellular volume.…”

Clathrin-coated structures support 3D directed migration through local force transmission

“…Following procedures previously described by Baschieri et al ( 25 ), control or siRNA-treated, genome-edited MDA-MB-231 cells were imaged for exposure times of 200 ms at 5-s intervals for the indicated time using a spinning disk microscope (Andor) based on a CSU-W1 Yokogawa head mounted on the lateral port of an inverted IX-83 Olympus microscope equipped with a 60× 1.35 NA UPLSAPO objective lens and a laser combiner system, which included 491- and 561-nm 100-mW DPSS lasers (Andor). Images were acquired with a Zyla sCMOS camera (Andor).…”