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Vol. 15, Issue 12, 5369-5382, December 2004
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* Graduate Division of Biological and Biomedical Sciences, Emory University, Atlanta, GA 30322;
Departments of Cell Biology, Emory University, Atlanta, GA 30322;
Departments of Dermatology, Emory University, Atlanta, GA 30322;
|| Center for Neurodegenerative Diseases, Emory University, Atlanta, GA 30322; and
Genentech, Inc., South San Francisco, CA 94080-4990
Submitted March 31, 2004;
Revised September 20, 2004;
Accepted September 21, 2004
Monitoring Editor: Keith Mostov
Cytoskeletal networks control organelle subcellular distribution and function. Herein, we describe a previously unsuspected association between intermediate filament proteins and the adaptor complex AP-3. AP-3 and intermediate filament proteins cosedimented and coimmunoprecipitated as a complex free of microtubule and actin binding proteins. Genetic perturbation of the intermediate filament cytoskeleton triggered changes in the subcellular distribution of the adaptor AP-3 and late endocytic/lysosome compartments. Concomitant with these architectural changes, and similarly to AP-3-null mocha cells, fibroblasts lacking vimentin were compromised in their vesicular zinc uptake, their organellar pH, and their total and surface content of AP-3 cargoes. However, the total content and surface levels, as well as the distribution of the transferrin receptor, a membrane protein whose sorting is AP-3 independent, remained unaltered in both AP-3- and vimentin-null cells. Based on the phenotypic convergence between AP-3 and vimentin deficiencies, we predicted and documented a reduced autophagosome content in mocha cells, a phenotype previously reported in cells with disrupted intermediate filament cytoskeletons. Our results reveal a novel role of the intermediate filament cytoskeleton in organelle/adaptor positioning and in regulation of the adaptor complex AP-3.
The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).
¶ Corresponding author. E-mail address: faundez{at}cellbio.emory.edu.
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