ATP binding regulates oligomerization and endosome association of RME-1 family proteins.
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.
Lee
Dong-won
DW
Laboratory of Cell Biology, NHLBI, National Institutes of Health, Bethesda, Maryland 20892-0301, USA.
Zhao
Xiaohong
X
Scarselletta
Sarah
S
Schweinsberg
Peter J
PJ
Eisenberg
Evan
E
Grant
Barth D
BD
Greene
Lois E
LE
D000251:Adenosine Triphosphatases / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry* / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D017353:Gene Deletion; D005982:Glutathione Transferase / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009154:Mutation; D009711:Nucleotides / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013997:Time Factors; D020798:Two-Hybrid System Techniques; D033921:Vesicular Transport Proteins / Q000737:chemistry / Q000378:metabolism*
D029742:Caenorhabditis elegans Proteins; C120700:EHD1 protein, human; C485640:Ehd1 protein, mouse; D009711:Nucleotides; C486205:RME-1 protein, C elegans; D033921:Vesicular Transport Proteins; D049452:Green Fluorescent Proteins; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D004247:DNA; D005982:Glutathione Transferase; D000251:Adenosine Triphosphatases