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MBC in Press, published online ahead of print June 7, 2006
Mol. Biol. Cell 10.1091/mbc.E05-10-0980

A more recent version of this article appeared on August 1, 2006
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Submitted on October 25, 2005
Revised on April 21, 2006
Accepted on May 25, 2006

Arf6-independent GEECs Fuse with Sorting Endosomes via a Rab5/PI-3' Kinase-dependent Machinery

Manjula Kalia,* Sudha Kumari,* Rahul Chadda,* Michelle M. Hill,{dagger} Robert G. Parton,{dagger} and Satyajit Mayor*

*National Centre for Biological Sciences, UAS-GKVK Campus, Bangalore 560065, India; {dagger}Institute for Molecular Bioscience, University of Queensland, Queensland 4072, Australia

Monitoring Editor: Jean Gruenberg

In the process of internalization of molecules from the extracellular milieu a cell utilizes multiple endocytic pathways, consequently generating different endocytic vesicles. These primary endocytic vesicles are targeted to specific destinations inside the cell. Here we show that GPI-anchored proteins are internalized by an Arf6-independent mechanism into GEECs (GPI-anchored protein-enriched early endosomal compartments). Internalized GPI-anchored proteins and the fluid phase are first visualized in GEECs that are acidic, primary endocytic structures, negative for early endosomal markers, Rab4, Rab5 and EEA1. They subsequently acquire Rab5 and EEA1 before homotypic fusion with other GEECs, and heterotypic fusion with endosomes containing cargo from the clathrin-dependent endocytic pathway. Although, the formation of GEECs is unaffected by inhibition of Rab5 GTPase and PI-3' kinase activity, their fusion with sorting endosomes is dependent on both activities. Over expression of Rab5 reverts PI-3' kinase inhibition of fusion, providing evidence that Rab5 effectors play important roles in heterotypic fusion between the dynamin-independent GEECs and clathrin and dynamin-dependent sorting endosomes.

Address correspondence to: Satyajit Mayor (mayor{at}ncbs.res.in)




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