1. Max Planck Institute of Molecular Cell Biology and Genetics,
2. MPI-CBG High-Throughput Technology Development Studio (HT-TDS), and
3. Scionics Computer Innovation GmbH, Pfotenhauerstrasse 108, 01307 Dresden, Germany
4. Cenix Bioscience GmbH, Tatzberg 47, 01307 Dresden, Germany

Correspondence to: Marino Zerial¹ Correspondence and requests for materials should be addressed to M.Z. (Email: zerial@mpi-cbg.de).

Abstract

Endocytosis is a key cellular process, encompassing different entry routes and endocytic compartments. To what extent endocytosis is subjected to high-order regulation by the cellular signalling machinery remains unclear. Using high-throughput RNA interference and automated image analysis, we explored the function of human kinases in two principal types of endocytosis: clathrin- and caveolae/raft-mediated endocytosis. We monitored this through infection of vesicular stomatitis virus, simian virus 40 and transferrin trafficking, and also through cell proliferation and apoptosis assays. Here we show that a high number of kinases are involved in endocytosis, and that each endocytic route is regulated by a specific kinase subset. Notably, one group of kinases exerted opposite effects on the two endocytic routes, suggesting coordinate regulation. Our analysis demonstrates that signalling functions such as those controlling cell adhesion, growth and proliferation, are built into the machinery of endocytosis to a much higher degree than previously recognized.

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