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PMID:17825285

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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

Significance

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

Notes

See also

References

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