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CALM Regulates Clathrin-Coated Vesicle Size and Maturation by Directly Sensing and Driving Membrane Curvature

DOI: 10.1016/j.devcel.2015.03.002 DOI Help

Authors: Sharon Miller (University of Cambridge) , Signe Mathiasen (University of Copenhagen) , Nicholas a. Bright (University of Cambridge) , Fabienne Pierre (CNRS) , Bernie Kelly (University of Cambridge) , Nikolay Kladt (Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases) , Astrid Schauss (Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases) , Christien j Merrifield (CNRS) , Dimitrios Stamou (University of Copenhagen) , Stefan Höning (University of Cologne) , David Owen (University of Cambridge)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Developmental Cell , VOL 33 , PAGES 163 - 175

State: Published (Approved)
Published: April 2015

Open Access Open Access

Abstract: The size of endocytic clathrin-coated vesicles (CCVs) is remarkably uniform, suggesting that it is optimized to achieve the appropriate levels of cargo and lipid internalization. The three most abundant proteins in mammalian endocytic CCVs are clathrin and the two cargo-selecting, clathrin adaptors, CALM and AP2. Here we demonstrate that depletion of CALM causes a substantial increase in the ratio of ‘‘open’’ clathrin-coated pits (CCPs) to ‘‘necked’’/ ‘‘closed’’ CCVs and a doubling of CCP/CCV diameter, whereas AP2 depletion has opposite effects. Depletion of either adaptor, however, significantly inhibits endocytosis of transferrin and epidermal growth factor. The phenotypic effects of CALM depletion can be rescued by re-expression of wildtype CALM, but not with CALM that lacks a functional N-terminal, membrane-inserting, curvature-sensing/ driving amphipathic helix, the existence and properties of which are demonstrated. CALM is thus a major factor in controlling CCV size and maturation and hence in determining the rates of endocytic cargo uptake. INTRODUC

Subject Areas: Biology and Bio-materials


Instruments: I03-Macromolecular Crystallography

Added On: 12/01/2016 13:36

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