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PMID: 9524117 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Distinct Rab-binding domains mediate the interaction of Rabaptin-5 with GTP-bound Rab4 and Rab5.

The EMBO journal ·Vol. 17 ·No. 7 ·1998-04-01 ·Pages 1941-51

Vitale G, Rybin V, Christoforidis S, Thornqvist P, McCaffrey M, Stenmark H, Zerial M

Abstract

Rabaptin-5 functions as an effector for the small GTPase Rab5, a regulator of endocytosis and early endosome fusion. We have searched for structural determinants that confer functional specificity on Rabaptin-5. Here we report that native cytosolic Rabaptin-5 is present in a homodimeric state and dimerization depends upon the presence of its coiled-coil predicted sequences. A 73 residue C-terminal region of Rabaptin-5 is necessary and sufficient both for the interaction with Rab5 and for Rab5-dependent recruitment of the protein on early endosomes. Surprisingly, we uncovered the presence of an additional Rab-binding domain at the N-terminus of Rabaptin-5. This domain mediates the direct interaction with the GTP-bound form of Rab4, a small GTPase that has been implicated in recycling from early endosomes to the cell surface. Based on these results, we propose that Rabaptin-5 functions as a molecular linker between two sequentially acting GTPases to coordinate endocytic and recycling traffic.

MeSH Terms
Amino Acid Sequence Animals Cattle Cell Line Cricetinae Cytosol/chemistry Dimerization Endosomes/metabolism GTP-Binding Proteins/analysis,metabolism Guanosine Triphosphate/metabolism HeLa Cells Humans Membrane Proteins/analysis,chemistry,genetics,metabolism Molecular Sequence Data Protein Binding Protein Structure, Secondary Recombinant Fusion Proteins Sequence Alignment Vesicular Transport Proteins rab4 GTP-Binding Proteins rab5 GTP-Binding Proteins
Chemicals
Membrane Proteins RABEP1 protein, human Recombinant Fusion Proteins Vesicular Transport Proteins Guanosine Triphosphate GTP-Binding Proteins rab4 GTP-Binding Proteins rab5 GTP-Binding Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Vitale G
European Molecular Biology Laboratory, Postfach 10.2209, D-69012 Heidelberg, Germany.
Rybin V
Christoforidis S
Thornqvist P
McCaffrey M
Stenmark H
Zerial M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.7.1941
Published
1998-04-01
Pages
1941-51
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170540
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/17.7.1941 PMC Full Text View journal details
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