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SH3-domain-containing proteins function at distinct steps in clathrin-coated vesicle formation
F Simpson1, N K Hussain, B Qualmann
1Department of Cell Biology, Scripps Research Institute, La Jolla, California 92037, USA.
Nature Cell Biology
|November 13, 1999
Summary
Several SH3 domains regulate endocytosis by interacting with dynamin. Specific SH3 domains, like intersectin
Area of Science:
- Cell biology
- Molecular biology
- Protein interactions
Background:
- SH3-domain-containing proteins are linked to endocytosis via dynamin interactions.
- The precise roles of these SH3 proteins in endocytosis are not fully understood.
Purpose of the Study:
- To investigate the function of SH3 domains in the formation of clathrin-coated vesicles.
- To elucidate the sequential roles of SH3 domain interactions in endocytic pathways.
Main Methods:
- Reconstitution of clathrin-coated vesicle formation in permeabilized 3T3-L1 cells.
- Assay of SH3 domain inhibition of vesicle formation in vitro.
- Analysis of SH3 domain interactions with membrane-associated proteins.
Main Results:
- SH3 domains of intersectin, endophilin I, syndapin I, and amphiphysin II inhibit vesicle formation.
- Most SH3 domains inhibit late-stage membrane fission events.
- The SH3A domain of intersectin specifically inhibits intermediate events, including constricted coated pit formation.
Conclusions:
- SH3 domain interactions with partners play sequential roles in endocytic coated-vesicle formation.
- Distinct SH3 domains regulate specific stages of the endocytic process.