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MBC in Press, published online ahead of print September 1, 2004
Mol. Biol. Cell 10.1091/mbc.E04-02-0127

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Submitted on February 13, 2004
Revised on August 20, 2004
Accepted on August 23, 2004

Protein Kinase C{delta} - Calmodulin Crosstalk Regulates Epidermal Growth Factor Receptor Exit from Early Endosomes

Anna Lladó,*{dagger} Francesc Tebar,*{dagger} Maria Calvo,* Jemina Moretó,* Alexander Sorkin,{ddagger} and Carlos Enrich{sect}

*Departament de Biologia Cellular, Facultat de Medicina, Universitat de Barcelona, Casanova 143, 08036 Barcelona, Spain; {ddagger}Department of Pharmacology, University of Colorado, Health Science Center, Denver, CO 80262

Monitoring Editor: Keith Mostov

We have recently shown that calmodulin antagonist (W13) interferes with the trafficking of the epidermal growth factor receptor (EGFR) and regulates the MAPK signaling pathway. In the present study we demonstrate that in cells in which calmodulin is inhibited, protein kinase C (PKC) inhibitors rapidly restores EGFR and transferrin trafficking through the recycling compartment, although onward transport to the degradative pathway remains arrested. Analysis of PKC isoforms reveals that inhibition of PKC{delta} with rottlerin or its down-modulation using small interfering RNA (siRNA) is specifically responsible for the release of the W13 blockage of EGFR trafficking from early endosomes. The use of the inhibitor Gö6976, specific for conventional PKCs ({alpha}, {beta}, {gamma}), or expression of dominant-negative forms of PKC{lambda}, {zeta} - or {epsilon} did not restore the effects of W13. Furthermore, in cells treated with W13 and rottlerin, a specific inhibitor of PKC{delta}, we observed a recovery of brefeldin A-tubulation, as well as transport of dextran-FITC toward the late endocytic compartment. These results demonstrate a specific interplay between calmodulin and PKC{delta} in the regulation of the morphology of and trafficking from the early endocytic compartment.

{dagger}These authors contributed equally to this work.

{sect}Corresponding author. E-mail: enrich{at}ub.edu




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