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PMID:20164305

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Citation

Tabuchi, M, Yanatori, I, Kawai, Y and Kishi, F (2010) Retromer-mediated direct sorting is required for proper endosomal recycling of the mammalian iron transporter DMT1. J. Cell. Sci. 123:756-66

Abstract

Endosomal recycling of the mammalian iron transporter DMT1 is assumed to be important for efficient and rapid uptake of iron across the endosomal membrane in the transferrin cycle. Here, we show that the retromer, a complex that mediates retrograde transport of transmembrane cargoes from endosomes to the trans-Golgi network, is required for endosomal recycling of DMT1-II, an alternative splicing isoform of DMT1. Bacterially expressed Vps26-Vsp29-Vsp35 trimer, a retromer cargo recognition complex, specifically binds to the cytoplasmic tail domain of DMT1-II in vitro. In particular, this binding is dependent on a specific hydrophobic motif of DMT1-II, which is required for its endosomal recycling. DMT1-II colocalizes with the Vps35 subunit of the retromer in TfR-positive endosomes. Depletion of the retromer by siRNA against Vps35 leads to mis-sorting of DMT1-II to LAMP2-positive structures, and expression of siRNA-resistant Vps35 can rescue this effect. These findings demonstrate that the retromer recognizes the recycling signal of DMT1-II and ensures its proper endosomal recycling.

Links

PubMed Online version:10.1242/jcs.060574

Keywords

Amino Acid Motifs/genetics; Amino Acid Motifs/physiology; Amino Acid Sequence; Cation Transport Proteins/chemistry; Cation Transport Proteins/genetics; Cation Transport Proteins/metabolism; Cell Line; Endosomes/metabolism; Humans; Microscopy, Fluorescence; Models, Biological; Molecular Sequence Data; Protein Transport/genetics; Protein Transport/physiology; RNA Interference; Sequence Homology, Amino Acid; Vesicular Transport Proteins/chemistry; Vesicular Transport Proteins/genetics; Vesicular Transport Proteins/metabolism; trans-Golgi Network/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:VPS35

located_in

GO:0005769: early endosome

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:VPS35

involved_in

GO:0032456: endocytic recycling

ECO:0000315: mutant phenotype evidence used in manual assertion

P

  • has_input:(UniProtKB:P49281-1)

Seeded From UniProt

complete

HUMAN:VPS35

part_of

GO:0030906: retromer, cargo-selective complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:NRAM2

enables

GO:1905394: retromer complex binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:Q8IUD7

enables

GO:1905394: retromer complex binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

Notes

See also

References

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