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PMID:15710626

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Citation

Lee, DW, Zhao, X, Scarselletta, S, Schweinsberg, PJ, Eisenberg, E, Grant, BD and Greene, LE (2005) ATP binding regulates oligomerization and endosome association of RME-1 family proteins. J. Biol. Chem. 280:17213-20

Abstract

Members of the RME-1/mRme-1/EHD1 protein family have recently been shown to function in the recycling of membrane proteins from recycling endosomes to the plasma membrane. RME-1 family proteins are normally found in close association with recycling endosomes and the vesicles and tubules emanating from these endosomes, consistent with the proposal that these proteins directly participate in endosomal transport. RME-1 family proteins contain a C-terminal EH (eps15 homology) domain thought to be involved in linking RME-1 to other endocytic proteins, a coiled-coil domain thought to be involved in homo-oligomerization and an N-terminal P-loop domain thought to mediate nucleotide binding. In the present study, we show that both Caenorhabditis elegans and mouse RME-1 proteins bind and hydrolyze ATP. No significant GTP binding or hydrolysis was detected. Mutation or deletion of the ATP-binding P-loop prevented RME-1 oligomerization and at the same time dissociated RME-1 from endosomes. In addition, ATP depletion caused RME-1 to lose its endosome association in the cell, resulting in cytosolic localization. Taken together, these results indicate that ATP binding is required for oligomerization of mRme-1/EHD1, which in turn is required for its association with endosomes.

Links

PubMed Online version:10.1074/jbc.M412751200

Keywords

Adenosine Triphosphatases/chemistry; Adenosine Triphosphate/chemistry; Adenosine Triphosphate/metabolism; Animals; Blotting, Western; Caenorhabditis elegans; Caenorhabditis elegans Proteins/chemistry; Caenorhabditis elegans Proteins/metabolism; Cell Line; Cell Membrane/metabolism; Cytosol/metabolism; DNA/metabolism; Endosomes/metabolism; Escherichia coli/metabolism; Gene Deletion; Glutathione Transferase/metabolism; Green Fluorescent Proteins/chemistry; Green Fluorescent Proteins/metabolism; Guanosine Triphosphate/chemistry; Guanosine Triphosphate/metabolism; HeLa Cells; Humans; Hydrolysis; Kinetics; Mice; Microscopy, Confocal; Microscopy, Fluorescence; Mutation; Nucleotides/chemistry; Protein Binding; Protein Structure, Tertiary; Time Factors; Two-Hybrid System Techniques; Vesicular Transport Proteins/chemistry; Vesicular Transport Proteins/metabolism

Significance

Annotations

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

CAEEL:G5ECC3

enables

GO:0016887: ATP hydrolysis activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CAEEL:G5EEU7

enables

GO:0016887: ATP hydrolysis activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CAEEL:G5EFP1

enables

GO:0016887: ATP hydrolysis activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CAEEL:Q86S80

enables

GO:0016887: ATP hydrolysis activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CAEEL:Q8WSP1

enables

GO:0016887: ATP hydrolysis activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CAEEL:Q9N5B7

enables

GO:0016887: ATP hydrolysis activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete


See also

References

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