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PMID: 11532964 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Hrs recruits clathrin to early endosomes.

The EMBO journal ·Vol. 20 ·No. 17 ·2001-09-03 ·Pages 5008-21

Raiborg C, Bache KG, Mehlum A, Stang E, Stenmark H

Abstract

The hepatocyte growth factor-regulated tyrosine kinase substrate, Hrs, has been implicated in intracellular trafficking and signal transduction. Hrs contains a phosphatidylinositol 3-phosphate-binding FYVE domain that contributes to its endosomal targeting. Here we show that Hrs and EEA1, a FYVE domain protein involved in endocytic membrane fusion, are localized to different regions of early endosomes. We demonstrate that Hrs co-localizes with clathrin, and that the C-terminus of Hrs contains a functional clathrin box motif that interacts directly with the terminal beta-propeller domain of clathrin heavy chain. A massive recruitment of clathrin to early endosomes was observed in cells transfected with Hrs, but not with Hrs lacking the C-terminus. Furthermore, the phosphatidylinositol 3-kinase inhibitor wortmannin caused the dissociation of both Hrs and clathrin from endosomes. While overexpression of Hrs did not affect endocytosis and recycling of transferrin, endocytosed epidermal growth factor and dextran were retained in early endosomes. These results provide a molecular mechanism for the recruitment of clathrin onto early endosomes and suggest a function for Hrs in trafficking from early to late endosomes.

MeSH Terms
Amino Acid Sequence Animals Binding Sites Cell Line Clathrin/chemistry,metabolism,physiology Cricetinae Dextrans/metabolism Endocytosis Endosomal Sorting Complexes Required for Transport Endosomes/physiology,ultrastructure Epidermal Growth Factor/metabolism Macromolecular Substances Melanoma Phosphatidylinositol Phosphates/metabolism Phosphoproteins/chemistry,metabolism,physiology Protein Conformation Protein Subunits Protein Transport Sequence Alignment Sequence Homology, Amino Acid Transferrin/metabolism Tumor Cells, Cultured
Chemicals
Clathrin Dextrans Endosomal Sorting Complexes Required for Transport Macromolecular Substances Phosphatidylinositol Phosphates Phosphoproteins Protein Subunits Transferrin hepatocyte growth factor-regulated tyrosine kinase substrate phosphatidylinositol 3-phosphate Epidermal Growth Factor
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Raiborg C
Department of Biochemistry, Institute for Cancer Research, The Norwegian Radium Hospital, Montebello, N-0310 Oslo, Norway.
Bache K G
Mehlum A
Stang E
Stenmark H
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.17.5008
Published
2001-09-03
Pages
5008-21
Language
English
Region
England
NLM ID
8208664
PMCID
PMC125612
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.17.5008 PMC Full Text View journal details
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