Clathrin-independent carriers form a high capacity endocytic sorting system at the leading edge of migrating cells

Howes, MT; Kirkham, M; Riches, J; Cortese, K; Walser, PJ; Simpson, F; Hill, MM; Jones, A; Lundmark, R; Lindsay, MR; Hernandez-Deviez, DJ; Hadzic, G; McCluskey, A; Bashir, R; Liu, L; Pilch, P; McMahon, H; Robinson, PJ; Hancock, JF; Mayor, S; Parton, RG.

doi:10.1083/jcb.201002119

Clathrin-independent carriers form a high capacity endocytic sorting system at the leading edge of migrating cells

Howes, MT; Kirkham, M; Riches, J; Cortese, K; Walser, PJ; Simpson, F; Hill, MM; Jones, A; Lundmark, R; Lindsay, MR; Hernandez-Deviez, DJ; Hadzic, G; McCluskey, A; Bashir, R; Liu, L; Pilch, P; McMahon, H; Robinson, PJ; Hancock, JF; Mayor, S; Parton, RG.

Authors

MT Howes

M Kirkham

J Riches

K Cortese

PJ Walser

F Simpson

MM Hill

A Jones

R Lundmark

MR Lindsay

DJ Hernandez-Deviez

G Hadzic

A McCluskey

R Bashir

L Liu

P Pilch

H McMahon

PJ Robinson

JF Hancock

S Mayor

RG. Parton

Abstract

Although the importance of clathrin- and caveolin-independent endocytic pathways has recently emerged, key aspects of these routes remain unknown. Using quantitative ultrastructural approaches, we show that clathrin-independent carriers (CLICs) account for approximately three times the volume internalized by the clathrin-mediated endocytic pathway, forming the major pathway involved in uptake of fluid and bulk membrane in fibroblasts. Electron tomographic analysis of the 3D morphology of the earliest carriers shows that they are multidomain organelles that form a complex sorting station as they mature. Proteomic analysis provides direct links between CLICs, cellular adhesion turnover, and migration. Consistent with this, CLIC-mediated endocytosis of key cargo proteins, CD44 and Thy-1, is polarized at the leading edge of migrating fibroblasts, while transient ablation of CLICs impairs their ability to migrate. These studies provide the first quantitative ultrastructural analysis and molecular characterization of the major endocytic pathway in fibroblasts, a pathway that provides rapid membrane turnover at the leading edge of migrating cells.

Citation

Howes, M., Kirkham, M., Riches, J., Cortese, K., Walser, P., Simpson, F., …Parton, R. (2010). Clathrin-independent carriers form a high capacity endocytic sorting system at the leading edge of migrating cells. Journal of Cell Biology, 190(4), 675-691. https://doi.org/10.1083/jcb.201002119

Journal Article Type	Article
Publication Date	Aug 1, 2010
Deposit Date	Apr 16, 2012
Publicly Available Date	Jun 12, 2012
Journal	Journal of Cell Biology
Print ISSN	0021-9525
Electronic ISSN	1540-8140
Publisher	Rockefeller University Press
Peer Reviewed	Peer Reviewed
Volume	190
Issue	4
Pages	675-691
DOI	https://doi.org/10.1083/jcb.201002119

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