Cell Calcium. 2008 May;43(5):432-43. Epub 2007 Sep 11.

RNF24, a new TRPC interacting protein, causes the intracellular retention of TRPC.External

Lussier, M. P., Lepage, P. K., Bousquet, S. M., Boulay, G.,
["Department of Pharmacology, Universite de Sherbrooke, Sherbrooke, Quebec, Canada J1H 5N4."]
TRPCs function as cation channels in non-excitable cells. The N-terminal tails of all TRPCs contain an ankyrin-like repeat domain, one of the most common protein-protein interaction motifs. Using a yeast two-hybrid screening approach, we found that RNF24, a new membrane RING-H2 protein, interacted with the ankyrin-like repeat domain of TRPC6. GST pull-down and co-immunoprecipitation assays showed that RNF24 interacted with all TRPCs. Cell surface-labelling assays showed that the expression of TRPC6 at the surface of HEK 293T cells was greatly reduced when it was transiently co-transfected with RNF24. Confocal microscopy showed that TRPC3 and TRPC6 co-localized with RNF24 in a perinuclear compartment and that RNF24 co-localized with mannosidase II, a marker of the Golgi cisternae. Using a pulse-chase approach, we showed that RNF24 did not alter the maturation process of TRPC6. Moreover, in HEK 293T cells, RNF24 did not alter carbachol-induced Ca(2+) entry via endogenous channels or TRPC6. These results indicate that RNF24 interacts with TRPCs in the Golgi apparatus and affects TRPC intracellular trafficking without affecting their activity.
PMID: 17850865External
Arrow2 Screening Arrow2 Validation: In vitro validation Arrow2 Validation: In vivo validation Arrow2 Characterization Arrow2 Functional consequence Arrow2 top
Screening Toggle
  Experimental screening Non-experimental screening Reference
TRP channel construct Interactor source
TRP channel Interactor Method Species Region Species Organ/tissue Sample type
TRPC1 Link RNF24 Inference Prediction 17850865
TRPC3 Link RNF24 Inference Prediction 17850865
TRPC4 Link RNF24 Inference Prediction 17850865
TRPC5 Link RNF24 Inference Prediction 17850865
TRPC6 Link RNF24 Yeast two-hybrid Not specified 3-259 Human Fetal brain cDNA library 17850865
TRPC7 Link RNF24 Inference Prediction 17850865
(Link: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
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Validation: In vitro validation Toggle
  Assay with recombinant proteins Reference
TRP channel construct Interactor construct
TRP channel Interactor Method Species Region Expression system Species Region Expression system
TRPC1 Link RNF24 Fusion protein-pull down assay Not specified 3-200 In vitro translation Human 98-148 E. coli 17850865
TRPC3 Link RNF24 Fusion protein-pull down assay Not specified 1-201 In vitro translation Human 98-148 E. coli 17850865
TRPC4 Link RNF24 Fusion protein-pull down assay Not specified 3-183 In vitro translation Human 98-148 E. coli 17850865
TRPC5 Link RNF24 Fusion protein-pull down assay Not specified 1-183 In vitro translation Human 98-148 E. coli 17850865
TRPC6 Link RNF24 Yeast two-hybrid Not specified 3-259 Yeast Human 98-148 Yeast 17850865
TRPC6 Link RNF24 Fusion protein-pull down assay Not specified 3-259 In vitro translation Human 98-148 E. coli 17850865
TRPC7 Link RNF24 Fusion protein-pull down assay Not specified 3-195 In vitro translation Human 98-148 E. coli 17850865
(Link: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
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Validation: In vivo validation Toggle
  Assay with endogenous proteins Assay with overexpressed proteins Reference
Cell or tissue Cell or tissue TRP channel construct Interactor construct
TRP channel Interactor Method Species Region Species Region
TRPC3 Link RNF24 Co-immunofluorescence staining HEK293 Not specified Full-length Human Full-length 17850865
TRPC6 Link RNF24 Co-immunofluorescence staining HEK293 Not specified Full-length Human Full-length 17850865
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Characterization Toggle
  Binding region mapping Stoichiometry Affinity (Kd) Reference
TRP channel Interactor
TRP channel Interactor Method Species Region Species Region
TRPC6 Link RNF24 Yeast two-hybrid Not specified 3-130 Human 98-148 17850865
TRPC6 Link RNF24 Yeast two-hybrid Not specified 122-259 Human 98-148 17850865
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Functional consequence Toggle
TRP channel Interactor Method Post-translational modification Subcellular trafficking Activity Reference
TRPC3 Link RNF24 Cell surface biotinylation Decrease in plasma membrane level 17850865
TRPC4 Link RNF24 Cell surface biotinylation Decrease in plasma membrane level 17850865
TRPC6 Link RNF24 Cell surface biotinylation Decrease in plasma membrane level 17850865
(Link: click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
TRP / Interactor

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