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MBC in Press, published online ahead of print September 29, 2004
Mol. Biol. Cell 10.1091/mbc.E04-03-0272

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Submitted on March 31, 2004
Revised on September 20, 2004
Accepted on September 21, 2004

The Endo-Lysosomal Sorting Machinery Interacts with the Intermediate Filament Cytoskeleton

Melanie L. Styers,*{dagger} Gloria Salazar,{dagger} Rachal Love,{dagger} Andrew A. Peden,{ddagger} Andrew P. Kowalczyk,{sect} and Victor Faundez{dagger}||¶

*Graduate Division of Biological and Biomedical Sciences, Departments of {dagger}Cell Biology and {sect}Dermatology, ||Center for Neurodegenerative Diseases, Emory University, Atlanta, GA 30322; and {ddagger}Genentech, Inc., South San Franscisco, CA 94080-4990

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.

Corresponding author. E-mail: faundez{at}cellbio.emory.edu




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