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Divalent Rab effectors regulate the sub-compartmental organization and sorting of early endosomes

Nature Cell Biology volume 4, pages 124–133 (2002)Cite this article

An Author Correction to this article was published on 05 December 2022

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Abstract

The three GTPases Rab5, Rab4 and Rab11 regulate sequential transport steps along the endocytic/recycling pathway, and occupy distinct membrane domains on early and recycling endosomes. To address the mechanisms that regulate communication between such domains, we searched for proteins that interact with both Rab5 and Rab4. Here, we report that Rabenosyn-5, a previously identified Rab5 effector, also binds to Rab4. Rabenosyn-5 overexpression increased the association between Rab5 and Rab4 endosomal domains and decreased the fraction of Rab4- and Rab11-positive structures. This redistribution was accompanied by a faster rate of transferrin recycling from early endosomes to the cell surface and reduced transport to Rab11-containing perinuclear recycling endosomes. These effects depend on the ability of Rabenosyn-5 to interact with Rab4. We propose that divalent Rab effectors regulate protein sorting and recycling by connecting Rab domains on early endosomes.

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Figure 1: Purification and identification of common effectors of Rab5 and Rab4 by sequential affinity chromatography.
Figure 2: Rabenosyn-5 interacts directly with Rab4 and recruits hVps45 to Rab4.
Figure 3: Confocal immunofluorescence analysis of Rab5-, Rab4- and Rabenosyn-5-labelled endosomes.
Figure 4: Rabenosyn-5 binds simultaneously to Rab5 and Rab4.
Figure 5: Overexpression of Rabenosyn-5 and Rabaptin-5, but not EEA1, increases Rab5 and Rab4 colocalization.
Figure 6: Rabenosyn-5 overexpression does not change the perinuclear localization of Rab11-positive membranes.
Figure 7: Morphological analysis of transferrin transport.
Figure 8: Effects of Rabenosyn-5 overexpression on transferrin recycling and EGF degradation.

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Acknowledgements

We are grateful to E. Nielsen, for many constructive comments and advice at the beginning of the project, and for anti-Rabenosyn-5 antibodies. We also thank A. Runge for technical assistance and B. Hoflack, H. McBride and K. Simons for critical reading of the manuscript and helpful comments. This work was supported by the Max Planck Society and by grants from Human Frontier Science Program (RG-0260/1999), European Union 5th Framework Programme (HPRN-CT-2000-00081) and Biomed (BMH4-97-2410) to M.Z.

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  1. Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse, 01307, Dresden, Germany

    Stefano De Renzis & Marino Zerial

  2. Cenix BioScience GmbH, Pfotenhauerstrasse, 01307, Dresden, Germany

    Birte Sönnichsen

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Correspondence to Marino Zerial.

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De Renzis, S., Sönnichsen, B. & Zerial, M. Divalent Rab effectors regulate the sub-compartmental organization and sorting of early endosomes. Nat Cell Biol 4, 124–133 (2002). https://doi.org/10.1038/ncb744

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