Article abstract
Nature Cell Biology 1, 119 - 124 (1999)
Published online: 15 May 1999 | doi:10.1038/10091
SH3-domain-containing proteins function at distinct steps in clathrin-coated vesicle formation
Fiona Simpson1, Natasha K. Hussain2, Britta Qualmann3, Regis B. Kelly3, Brian K. Kay4, Peter S. McPherson2 & Sandra L. Schmid1
Abstract
Several SH3-domain-containing proteins have been implicated in endocytosis by virtue of their interactions with dynamin; however, their functions remain undefined. Here we report the efficient reconstitution of ATP-, GTP-, cytosol- and dynamin-dependent formation of clathrin-coated vesicles in permeabilized 3T3-L1 cells. The SH3 domains of intersectin, endophilin I, syndapin I and amphiphysin II inhibit coated-vesicle formation in vitro through interactions with membrane-associated proteins. Most of the SH3 domains tested selectively inhibit late events involving membrane fission, but the SH3A domain of intersectin uniquely inhibits intermediate events leading to the formation of constricted coated pits. These results suggest that interactions between SH3 domains and their partners function sequentially in endocytic coated-vesicle formation.
- Department of Cell Biology, The Scripps Research Institute , 10550 N. Torrey Pines, La Jolla, California 92037, USA
- Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Quebec H3A 2B4, Canada
- Department of Biochemistry and Biophysics and the Hormone Research Institute, University of California, San Francisco, California 94143-0534, USA
- Department of Pharmacology, University of Wisconsin , Madison, Wisconsin 53706-1532, USA
Correspondence to: Peter S. McPherson2 mcpm@musica.mcgill.ca
