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MBC in Press, published online ahead of print May 17, 2006
Mol. Biol. Cell 10.1091/mbc.E05-11-1054

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Submitted on November 16, 2005
Revised on April 28, 2006
Accepted on May 8, 2006

Distinct Roles for Tsg101 and Hrs in Multivesicular Body Formation and Inward Vesiculation

M. Razi and C. E. Futter

Institute of Ophthalmology, UCL, London EC1V 9EL, United Kingdom

Monitoring Editor: Sandra Schmid

In mammalian cells EGF stimulation promotes multivesicular body (MVB) formation and inward vesiculation within MVB. Annexin 1 is required for EGF-stimulated inward vesiculation but not MVB formation, demonstrating that MVB formation (the number of MVB/U cytoplasm) and inward vesiculation (the number of internal vesicles/MVB) are regulated by different mechanisms. Here we show that EGF-stimulated MVB formation requires Tsg101, a component of the ESCRT machinery. Depletion of Tsg101 potently inhibits EGF degradation and MVB formation and causes the vacuolar domains of the early endosome to tubulate. Although Tsg101 depletion inhibits MVB formation and alters the morphology of the early endosome in unstimulated cells, these effects are much greater following EGF stimulation. In contrast, depletion of Hrs only modestly inhibits EGF degradation, does not induce tubulation of the early endosome and causes the generation of enlarged MVBs that retain the ability to fuse with the lysosome. Taken together these results indicate that Tsg101 is required for the formation of stable vacuolar domains within the early endosome that develop into MVBs and Hrs is required for the accumulation of internal vesicles within MVBs and that both these processes are up-regulated by EGF stimulation.

Address correspondence to: C. E. Futter (c.futter{at}ucl.ac.uk)




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