PMID:17825285

Citation	Yang, Q, Onuki, R, Nakai, C and Sugiyama, Y (2007) Ezrin and radixin both regulate the apical membrane localization of ABCC2 (MRP2) in human intestinal epithelial Caco-2 cells. Exp. Cell Res. 313:3517-25
Abstract	Multidrug resistance-associated protein ABCC2 (MRP2) is widely expressed in mammalian tissues including intestine, liver and kidney, and it has been shown to be located exclusively on the apical membrane of polarized cells. Recently, several reports suggest that apical membrane localization of ABCC2 (Mrp2) was regulated by radixin in rodent liver. To investigate the mechanism underlying this apical membrane targeting of MRP2 in human intestine, we chose Caco-2 cells as a model to examine the unique roles of ezrin and radixin. Following immunostaining, radixin and ezrin were found to be concentrated at the apical membrane of Caco-2 cells. Using the RNAi method, radixin and ezrin stable knockdown Caco-2 cells were constructed. A cell surface biotinylation experiment with radixin or ezrin stable knockdown Caco-2 cells showed that radixin or ezrin deficiency caused the loss of ABCC2 (MRP2) from the cell surface. An immunoprecipitation assay showed that radixin and ezrin were associated with ABCC2 (MRP2). These findings indicate that both ezrin and radixin are independently required for the apical membrane localization of ABCC2 (MRP2) in Caco-2 cells. Radixin and ezrin play similar roles in the apical membrane localization of ABCC2 (MRP2) and their expression level and subcellular distribution are important factors in the regulation of ABCC2 (MRP2) at the post-transcriptional level.
Links	PubMed Online version:10.1016/j.yexcr.2007.07.033
Keywords	Actins/metabolism; Caco-2 Cells; Cell Membrane/metabolism; Cell Polarity; Cytoskeletal Proteins/metabolism; Epithelial Cells/cytology; Fluorescent Antibody Technique; Humans; Intestines/cytology; Membrane Proteins/metabolism; Membrane Transport Proteins/metabolism; Multidrug Resistance-Associated Proteins/metabolism; P-Glycoprotein/metabolism; Phosphoproteins/metabolism; Protein Binding; Protein Transport; RNA, Small Interfering/metabolism; Zonula Occludens-1 Protein
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
HUMAN:EZRI	located_in	GO:0016324: apical plasma membrane	ECO:0000314: direct assay evidence used in manual assertion		C	part_of:(CL:0002563)	Seeded From UniProt	complete
HUMAN:EZRI	involved_in	GO:0072659: protein localization to plasma membrane	ECO:0000315: mutant phenotype evidence used in manual assertion		P	has_input:(UniProtKB:Q92887) occurs_in:(CL:0002563) occurs_at:(GO:0016324)	Seeded From UniProt	complete
HUMAN:RADI	located_in	GO:0016324: apical plasma membrane	ECO:0000314: direct assay evidence used in manual assertion		C	part_of:(CL:0002563)	Seeded From UniProt	complete
HUMAN:RADI	enables	GO:0003779: actin binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
HUMAN:RADI	involved_in	GO:0072659: protein localization to plasma membrane	ECO:0000315: mutant phenotype evidence used in manual assertion		P	has_input:(UniProtKB:Q92887) occurs_in:(CL:0002563) occurs_at:(GO:0016324)	Seeded From UniProt	complete
HUMAN:RADI	enables	GO:0051117: ATPase binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:Q92887	F		Seeded From UniProt	complete
HUMAN:MRP2	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P15311	F		Seeded From UniProt	complete
HUMAN:MRP2	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P35241	F		Seeded From UniProt	complete
HUMAN:MRP2	located_in	GO:0009986: cell surface	ECO:0000314: direct assay evidence used in manual assertion		C	part_of:(CL:0002563)	Seeded From UniProt	complete
HUMAN:MRP2	located_in	GO:0016324: apical plasma membrane	ECO:0000314: direct assay evidence used in manual assertion		C	part_of:(CL:0002563)	Seeded From UniProt	complete
HUMAN:F5H1A7	part_of	GO:0016324: apical plasma membrane	ECO:0000314: direct assay evidence used in manual assertion		C	part_of:(CL:0002563)	Seeded From UniProt	complete
HUMAN:F5H1A7	enables	GO:0051117: ATPase binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:Q92887	F		Seeded From UniProt	complete
HUMAN:F5H1A7	enables	GO:0003779: actin binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
HUMAN:F5H1A7	involved_in	GO:0072659: protein localization to plasma membrane	ECO:0000315: mutant phenotype evidence used in manual assertion		P	has_direct_input:(UniProtKB:Q92887) occurs_in:(CL:0002563) occurs_at:(GO:0016324)	Seeded From UniProt	complete
HUMAN:EZRI	part_of	GO:0016324: apical plasma membrane	ECO:0000314: direct assay evidence used in manual assertion		C	part_of:(CL:0002563)	Seeded From UniProt	complete
HUMAN:EZRI	enables	GO:0003779: actin binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
HUMAN:EZRI	enables	GO:0051117: ATPase binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:Q92887	F		Seeded From UniProt	complete
HUMAN:EZRI	involved_in	GO:0072659: protein localization to plasma membrane	ECO:0000315: mutant phenotype evidence used in manual assertion		P	has_direct_input:(UniProtKB:Q92887) occurs_in:(CL:0002563) occurs_at:(GO:0016324)	Seeded From UniProt	complete

References

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