Mutations in human dynamin block an intermediate stage in coated vesicle formation
Abstract
The role of human dynamin in receptor-mediated endocytosis was investigated by transient expression of GTP-binding domain mutants in mammalian cells. Using assays which detect intermediates in coated vesicle formation, the dynamin mutants were found to block endocytosis at a stage after the initiation of coat assembly and preceding the sequestration of ligands into deeply invaginated coated pits. Membrane transport from the ER to the Golgi complex was unaffected indicating that dynamin mutants specifically block early events in endocytosis. These results demonstrate that mutations in the GTP-binding domain of dynamin block Tfn-endocytosis in mammalian cells and suggest that a functional dynamin GTPase is required for receptor-mediated endocytosis via clathrin-coated pits.
Additional Information
© 1993 The Rockefeller University Press Received for publication 15 April 1993 and in revised form 27 April 1993. The human dynamin cDNA sequences are accessible under GenBank accession numbers L07807-L07810. The first two authors contributed equally to this work. We world like to thank Drs. Reid Gilmore, Harris Berustein, and Peter Walter for helpful discussions and for communicating their results before publication, E. Koh for technical assistance and George Klier for assistance with confocal microscopy and photography. This work was supported by National Institutes of Health (NIH) grant GM42445 and a grant from the Lucille P. Markey Charitable Trust to S.L. Schmid, and an NIH Program Project grant GM40499 to E.M. Meyerowitz. S.L. Schmid is a Lucille P. Markey Scholar. E.J. Tisdale was supported by an NIH Postdoctoral Fellowship CA09270. A.M.v.d. Bliek was supported by a Long Term Fellowship from the Human Frontier Science Program. H. Damke was supported by a fellowship from the Deutsche Forschungsgemeinschaft. This is manuscript 7711-CB from The Scripps Research Institute.Attached Files
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Additional details
- PMCID
- PMC2119674
- Eprint ID
- 5143
- Resolver ID
- CaltechAUTHORS:BLIjcb93
- NIH
- GM42445
- Lucille P. Markey Charitable Trust
- NIH
- GM40499
- NIH Postdoctoral Fellowship
- CA09270
- Human Frontier Science Program
- Deutsche Forschungsgemeinschaft (DFG)
- Created
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2006-10-03Created from EPrint's datestamp field
- Updated
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2021-11-08Created from EPrint's last_modified field