Cargo regulates clathrin-coated pit invagination via clathrin light chain phosphorylation

Maib, Hannes; Ferreira, Filipe; Vassilopoulos, Stéphane; Smythe, Elizabeth

doi:10.1083/jcb.201805005

Cited by 26 publications

(28 citation statements)

References 80 publications

(127 reference statements)

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“…However, essentially all known functions ascribed to clathrin light chains appear to be shared by both light chains and, where identified, involve identical sequences. These include modulation of in vitro clathrin lattice assembly and disassembly (Brodsky, 2012;Schmid et al, 1984;Ungewickell and Ungewickell, 1991), LRRK2 binding and Rac1 inactivation (Schreij et al, 2015), gyrating-clathrin (G-clathrin) formation and cargo recycling (Luo et al, 2013;Majeed et al, 2014;Parachoniak et al, 2011;Zhao and Keen, 2008, and this study), internalization of some G-protein-coupled receptors (GPCRs) (Ferreira et al, 2012;Maib et al, 2018) and Hip1-mediated actin interaction (Chen and Brodsky, 2005;Engqvist-Goldstein et al, 2001;Legendre-Guillemin et al, 2002;Legendre-Guillemin et al, 2005). Recently, a role for CLCb in the modulation of endocytic coated pit dynamics and EGFR processing has been identified (Chen et al, 2017b), and the importance of CLCa for internalization of some GPCRs has been inferred from immunological phenotypes in knockout mice (Wu et al, 2016), validating the concept of CLC-specific functions.…”

Section: Introductionmentioning

confidence: 85%

A unique role for clathrin light chain a in cell spreading and migration

Tsygankova

Keen

2019

Journal of Cell Science

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Clathrin heavy chain is the structural component of the clathrin triskelion, but unique functions for the two distinct and highly conserved clathrin light chains (CLCa and CLCb, also known as CLTA and CLTB, respectively) have been elusive. Here, we show that following detachment and replating, CLCa is uniquely responsible for promoting efficient cell spreading and migration. Selective depletion of CLCa, but not of CLCb, reduced the initial phase of isotropic spreading of HeLa, H1299 and HEK293 cells by 60-80% compared to siRNA controls, and wound closure and motility by ∼50%. Surface levels of β1-integrins were unaffected by CLCa depletion. However, CLCa was required for effective targeting of FAK (also known as PTK2) and paxillin to the adherent surface of spreading cells, for integrin-mediated activation of Src, FAK and paxillin, and for maturation of focal adhesions, but not their microtubule-based turnover. Depletion of CLCa also blocked the interaction of clathrin with the nucleation-promoting factor WAVE complex, and altered actin distribution. Furthermore, preferential recruitment of CLCa to budding protrusions was also observed. These results comprise the first identification of CLCa-specific functions, with implications for normal and neoplastic integrin-based signaling and cell migration.

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Section: Introductionmentioning

confidence: 85%

A unique role for clathrin light chain a in cell spreading and migration

Tsygankova

Keen

2019

Journal of Cell Science

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“…Both P2Y 12 and μOR require phosphorylated clathrin light chain for efficient CME, whereas TfR does not. 80,81 B2AR endocytosis is blocked at 16 C while TfR endocytosis is not. 77 Segregation of different cargo into specific subsets of CCPs could allow individual control over clustering and endocytosis of different receptors.…”

Section: Segregation Of Gpcrs In Endocytic Domainsmentioning

confidence: 94%

“…GPCRs also show different biochemical requirements for endocytosis. Both P2Y 12 and μOR require phosphorylated clathrin light chain for efficient CME, whereas TfR does not . B2AR endocytosis is blocked at 16°C while TfR endocytosis is not .…”

Section: Agonist‐mediated Receptor Endocytosismentioning

confidence: 95%

Regulation of G protein‐coupled receptor signaling by plasma membrane organization and endocytosis

Weinberg

Puthenveedu

2019

Traffic

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The trafficking of G protein coupled‐receptors (GPCRs) is one of the most exciting areas in cell biology because of recent advances demonstrating that GPCR signaling is spatially encoded. GPCRs, acting in a diverse array of physiological systems, can have differential signaling consequences depending on their subcellular localization. At the plasma membrane, GPCR organization could fine‐tune the initial stages of receptor signaling by determining the magnitude of signaling and the type of effectors to which receptors can couple. This organization is mediated by the lipid composition of the plasma membrane, receptor‐receptor interactions, and receptor interactions with intracellular scaffolding proteins. GPCR organization is subsequently changed by ligand binding and the regulated endocytosis of these receptors. Activated GPCRs can modulate the dynamics of their own endocytosis through changing clathrin‐coated pit dynamics, and through the scaffolding adaptor protein β‐arrestin. This endocytic regulation has signaling consequences, predominantly through modulation of the MAPK cascade. This review explores what is known about receptor sorting at the plasma membrane, protein partners that control receptor endocytosis, and the ways in which receptor sorting at the plasma membrane regulates downstream trafficking and signaling.

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“…Limited functional differences between CLCa and CLCb have been observed in cell culture with respect to clathrin dynamics 23 , during focal adhesion formation 24 and during cancer cell migration 25 , with mechanisms attributed to possible isoform-specific differences in CLC binding proteins and post-translational modification 26 . The capacity for biophysical differences in clathrin comprising different CLC isoforms and their splicing variants to influence tissue-specific clathrin functions has not yet been considered.…”

Section: Introductionmentioning

confidence: 99%

“…Thus, CLCs affect clathrin-mediated membrane deformation directly through their influence on clathrin lattice properties 30,34 and indirectly through their interaction with the force-generating actin machinery. Loss of CLCs in culture and in vivo affects CCV uptake of some, but not all cargo, possibly reflecting variability in mechanical demand for packaging different cargo into CCVs 10,26,35 .…”

Section: Introductionmentioning

confidence: 99%

Clathrin light chain diversity regulates membrane deformation in vitro and synaptic vesicle formation in vivo

Redlingshoefer

McLeod

Camus

et al. 2019

Preprint

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Clathrin light chain (CLC) subunits in vertebrates are encoded by paralogous genes CLTA and CLTB and both gene products undergo alternative splicing in neurons. To understand how this CLC diversity influences neuronal clathrin function, we characterised the biophysical properties of clathrin comprising individual CLC variants for correlation with neuronal phenotypes of mice lacking either CLC-encoding gene. CLC variants differentially influenced clathrin knee conformation within assemblies, and clathrin lattices with neuronal CLC mixtures were more effective in membrane bending than those with single neuronal isoforms nCLCa or nCLCb. Correspondingly, electrophysiological recordings revealed that neurons from mice deficient for nCLCa or nCLCb were defective in synaptic vesicle recycling. Mice with only nCLCb had a reduced synaptic vesicle pool compared to wild-type mice, while nCLCa-only mice had increased synaptic vesicle numbers. These findings highlight functional differences between the CLC isoforms and show that isoform mixing influences tissue-specific clathrin function in neurons.

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Cargo regulates clathrin-coated pit invagination via clathrin light chain phosphorylation

Abstract: Phosphorylation of clathrin light chains (CLCs) regulates GPCR uptake but is dispensable for transferrin internalization. Maib et al. show that CLCb phosphorylation is required for efficient auxilin-mediated clathrin exchange to promote coated pit invagination in a cargo-specific manner.

Cited by 26 publications

References 80 publications

A unique role for clathrin light chain a in cell spreading and migration

A unique role for clathrin light chain a in cell spreading and migration

Regulation of G protein‐coupled receptor signaling by plasma membrane organization and endocytosis

Clathrin light chain diversity regulates membrane deformation in vitro and synaptic vesicle formation in vivo

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