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Vol. 12, Issue 9, 2790-2799, September 2001

Clathrin Hub Expression Affects Early Endosome Distribution with Minimal Impact on Receptor Sorting and Recycling

Elizabeth M. Bennett,*dagger Sharron X. Lin,Dagger Mhairi C. Towler,* Frederick R. Maxfield,Dagger and Frances M. Brodsky*dagger §||

 *The G. W. Hooper Foundation, Department of Microbiology and Immunology and Departments of  §Biopharmaceutical Sciences and  dagger Pharmaceutical Chemistry, University of California, San Francisco, California 94143-0552; and  Dagger Department of Biochemistry, Weill Medical College of Cornell University, New York, New York 10021

Clathrin-coated vesicles execute receptor-mediated endocytosis at the plasma membrane. However, a role for clathrin in later endocytic trafficking processes, such as receptor sorting and recycling or maintaining the organization of the endocytic pathway, has not been thoroughly characterized. The existence of clathrin-coated buds on endosomes suggests that clathrin might mediate later endocytic trafficking events. To investigate the function of clathrin-coated buds on endosomal membranes, endosome function and distribution were analyzed in a HeLa cell line that expresses the dominant-negative clathrin inhibitor Hub in an inducible manner. As expected, Hub expression reduced receptor-mediated endocytosis at the plasma membrane. Hub expression also induced a perinuclear aggregation of early endosome antigen 1-positive early endosomes, such that sorting and recycling endosomes were found tightly concentrated in the perinuclear region. Despite the dramatic redistribution of endosomes, Hub expression did not affect the overall kinetics of receptor sorting or recycling. These data show that clathrin function is necessary to maintain proper cellular distribution of early endosomes but does not play a prominent role in sorting and recycling events. Thus, clathrin's role on endosomal membranes is to influence organelle localization and is distinct from its role in trafficking pathways at the plasma membrane and trans-Golgi network.

|| Corresponding author. E-mail address: fmarbro{at}itsa.ucsf.edu.

Molecular Biology of the Cell
Vol. 12, 2790-2799, September 2001
Copyright © 2001 by The American Society for Cell Biology


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