Deciphering dynamics of clathrin-mediated endocytosis in a living organism

Ferguson, Joshua P.; Willy, Nathan M.; Heidotting, Spencer P.; Huber, Scott D.; Webber, Matthew J.; Kural, Cömert

doi:10.1083/jcb.201604128

Cited by 54 publications

(104 citation statements)

References 43 publications

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“…Clathrin transports a large variety of different cargoes from the plasma membrane in response to receptor-mediated endocytosis signals, and it is composed of a clathrin light chain and a clathrin heavy chain (Halebian et al, 2017). In clathrin-mediated endocytosis, vesicle scission is accelerated by the large GTPase dynamin, which involves the dynamic function of dynamin-2 at the neck of the clathrin-coated pit (Ferguson et al, 2016). Our results indicate that through transfecting siRNAs targeting CHC, we reduced clathrin expression in PK-15 cells without affecting PSV infection.…”

Section: Discussionmentioning

confidence: 99%

Porcine sapelovirus enters PK-15 cells via caveolae-dependent endocytosis and requires Rab7 and Rab11

Zhao

Cui

et al. 2019

Virology

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A B S T R A C TTo comprehensively understand the endocytosis of Sapelovirus A (PSV) entry into PK-15 cells, we studied PSV infection in the context of cell perturbations through drug inhibition, siRNA silencing and overexpression of dominant negative (DN) mutants. We showed here that PSV infection of PK-15 cells was unaffected by pretreated with chlorpromazine, EIPA, knockdown of the clathrin heavy chain or overexpression of Eps15 DN mutant. Conversely, PSV infection was sensitive to NH 4 Cl, chloroquine, dynasore, nystatin, MβCD and wortmannin with reduced PSV VP1 expression levels and virus titer. Additionally, PSV invasion leaded to rapid actin rearrangement and disruption of the cellular actin network enhanced PSV infection. After internalization the virus was transported to late endosomes and/or cycling endosomes that requires the participation of Rab7 and Rab11. Our findings demonstrate that PSV uses caveolae-dependent endocytosis as the predominant entry portal into PK-15 cells which requires low pH, dynamin, Rab7 and Rab11.

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Section: Discussionmentioning

confidence: 99%

Porcine sapelovirus enters PK-15 cells via caveolae-dependent endocytosis and requires Rab7 and Rab11

Zhao

Cui

et al. 2019

Virology

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“…Interestingly, CCPs are spatially organized in a migrating cell along the posterior and anterior axis a well as between ventral and dorsal surfaces . Furthermore, CCP dynamics slowed down during morphological changes in a Drosophila embryo, illustrating an effect of mechanical cues on endocytosis during development . In addition to the endocytic regulation of cell surface receptors that are involved in cell migration, in recent years, other membrane trafficking machineries have been connected to cell migration.…”

Section: Bringing It All Together: Modeling and Integration Into Systmentioning

confidence: 99%

The big and intricate dreams of little organelles: Embracing complexity in the study of membrane traffic

Liu

Botelho

Antonescu

2017

Traffic

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Compartmentalization of eukaryotic cells into dynamic organelles that exchange material through regulated membrane traffic governs virtually every aspect of cellular physiology including signal transduction, metabolism and transcription. Much has been revealed about the molecular mechanisms that control organelle dynamics and membrane traffic and how these processes are regulated by metabolic, physical and chemical cues. From this emerges the understanding of the integration of specific organellar phenomena within complex, multiscale and nonlinear regulatory networks. In this review, we discuss systematic approaches that revealed remarkable insight into the complexity of these phenomena, including the use of proximity-based proteomics, high-throughput imaging, transcriptomics and computational modeling. We discuss how these methods offer insights to further understand molecular versatility and organelle heterogeneity, phenomena that allow a single organelle population to serve a range of physiological functions. We also detail on how transcriptional circuits drive organelle adaptation, such that organelles may shift their function to better serve distinct differentiation and stress conditions. Thus, organelle dynamics and membrane traffic are functionally heterogeneous and adaptable processes that coordinate with higher-order system behavior to optimize cell function under a range of contexts. Obtaining a comprehensive understanding of organellar phenomena will increasingly require combined use of reductionist and system-based approaches.

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“…Clathrin-mediated endocytosis (CME) involves the internalization of cargo by sculpting plasma membrane into 60 -120 nm sized buds, supported by a clathrin protein coat 1 . CME is a well-studied endocytic pathway present in organisms at all developmental stages [1][2][3][4] . It plays a critical role in nutrient uptake, intracellular trafficking, and signal transduction 1,3 .…”

Section: Introductionmentioning

confidence: 99%

Complimentary action of structured and unstructured domains of epsin supports clathrin-mediated endocytosis at high tension

Joseph

Osoria

Yee

et al. 2020

Preprint

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Membrane tension plays an inhibitory role in clathrin-mediated endocytosis (CME) by impeding the transition of flat plasma membrane to hemispherical clathrin-coated structures (CCSs). Membrane tension also impedes the transition of hemispherical domes to omegashaped CCSs, a necessary step before their internalization via dynamin-mediated membrane scission. However, CME is not completely halted in cells under high tension conditions. Here we find that epsin, a membrane bending protein which inserts its N-terminus H0 helix into lipid bilayer, supports flat-to-dome transition and increases the stability of CCSs at high tension.This discovery is supported by molecular dynamic simulation of the epsin N-terminal homology (ENTH) domain that becomes more structured when embedded in a lipid bilayer. In addition, epsin has an intrinsically disordered protein (IDP) C-terminus domain which induces membrane curvature via steric repulsion. Insertion of H0 helix into lipid bilayer is not sufficient for stable epsin recruitment as deleting the IDP domain in epsin renders it cytosolic. Epsin's binding to adaptor protein 2 and clathrin is critical for epsin's association with CCSs under high tension conditions, supporting the importance of multivalent interactions in CCSs.Together, our results support a model where the ENTH and IDP domains of epsin have complementary roles to ensure CME initiation and CCS maturation are unimpeded under high tension environments.

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Deciphering dynamics of clathrin-mediated endocytosis in a living organism

Cited by 54 publications

References 43 publications

Porcine sapelovirus enters PK-15 cells via caveolae-dependent endocytosis and requires Rab7 and Rab11

Porcine sapelovirus enters PK-15 cells via caveolae-dependent endocytosis and requires Rab7 and Rab11

The big and intricate dreams of little organelles: Embracing complexity in the study of membrane traffic

Complimentary action of structured and unstructured domains of epsin supports clathrin-mediated endocytosis at high tension

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