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New tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision

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Wood, Laura A., Larocque, Gabrielle, Clarke, Nicholas I., Sarkar, Sourav and Royle, Stephen J. (2017) New tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision. The Journal of Cell Biology, 216 (12). 4351. doi:10.1083/jcb.201702188

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Official URL: http://dx.doi.org/10.1083/jcb.201702188

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Abstract

Clathrin-mediated endocytosis (CME) is the major route of receptor internalization at the plasma membrane. Analysis of constitutive CME is difficult because the initiation of endocytic events is unpredictable. When and where a clathrin-coated pit will form and what cargo it will contain are difficult to foresee. Here we describe a series of genetically encoded reporters that allow the initiation of CME on demand. A clathrin-binding protein fragment (“hook”) is inducibly attached to an “anchor” protein at the plasma membrane, which triggers the formation of new clathrin-coated vesicles. Our design incorporates temporal and spatial control by the use of chemical and optogenetic methods for inducing hook–anchor attachment. Moreover, the cargo is defined. Because several steps in vesicle creation are bypassed, we term it “hot-wiring.” We use hot-wired endocytosis to describe the functional interactions between clathrin and AP2. Two distinct sites on the β2 subunit, one on the hinge and the other on the appendage, are necessary and sufficient for functional clathrin engagement.

Item Type: Journal Article
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Faculty of Medicine > Warwick Medical School > Biomedical Sciences > Cell & Developmental Biology
Library of Congress Subject Headings (LCSH): Endocytosis, Cell membranes
Journal or Publication Title: The Journal of Cell Biology
Publisher: Rockefeller University Press
ISSN: 0021-9525
Official Date: 27 September 2017
Dates:
DateEvent
27 September 2017Available
25 August 2017Accepted
Volume: 216
Number: 12
Article Number: 4351
DOI: 10.1083/jcb.201702188
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Open Access
Funder: Medical Research Council (Great Britain) (MRC), Fonds québécois de la recherche sur la nature et les technologies [Quebec Fund for Nature and Technology Research] (FQRNT)
Grant number: MR/J003964/1 (MRC)
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
MR/J003964/1[MRC] Medical Research Councilhttp://dx.doi.org/10.13039/501100000265
UNSPECIFIEDFonds québécois de la recherche sur la nature et les technologies [Quebec Fund for Nature and Technology Research] (FQRNT)UNSPECIFIED

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