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New tools for "hot-wiring" clathrin-mediated endocytosis with temporal and spatial precision.
- Source :
-
The Journal of cell biology [J Cell Biol] 2017 Dec 04; Vol. 216 (12), pp. 4351-4365. Date of Electronic Publication: 2017 Sep 27. - Publication Year :
- 2017
-
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.<br /> (© 2017 Wood et al.)
- Subjects :
- Adaptor Protein Complex 2 metabolism
Cell Line
Clathrin metabolism
Clathrin-Coated Vesicles ultrastructure
Coated Pits, Cell-Membrane ultrastructure
Epithelial Cells ultrastructure
Gene Expression Regulation
Genes, Reporter
Green Fluorescent Proteins genetics
Green Fluorescent Proteins metabolism
HeLa Cells
Humans
Luminescent Proteins genetics
Luminescent Proteins metabolism
Metabolic Engineering methods
Optogenetics
Protein Binding
Protein Subunits genetics
Protein Subunits metabolism
Retinal Pigment Epithelium metabolism
Retinal Pigment Epithelium ultrastructure
Signal Transduction
Tacrolimus pharmacology
Tacrolimus Binding Proteins genetics
Tacrolimus Binding Proteins metabolism
Red Fluorescent Protein
Adaptor Protein Complex 2 genetics
Clathrin genetics
Clathrin-Coated Vesicles metabolism
Coated Pits, Cell-Membrane metabolism
Endocytosis genetics
Epithelial Cells metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1540-8140
- Volume :
- 216
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- The Journal of cell biology
- Publication Type :
- Academic Journal
- Accession number :
- 28954824
- Full Text :
- https://doi.org/10.1083/jcb.201702188