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TRPCs interact with RNF24 at the level of the Golgi apparatus
Author(s) -
Lussier Marc P.,
Lepage Pascale K.,
Boulay Guylain
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a245-b
Subject(s) - trpc3 , trpc , microbiology and biotechnology , trpv , golgi apparatus , transient receptor potential channel , chemistry , trpc6 , biophysics , endoplasmic reticulum , biochemistry , biology , receptor , trpv1
Transient receptor potential canonical channels (TRPCs) have been proposed to function as cation channels in non‐excitable cells. Amino acid sequence analysis of TRPCs reveals the presence of an ankyrin‐like repeat domain (ARD) in the N‐terminal tail. Using a yeast two‐hybrid screening, we found that RNF24, a new C3H2C3 RING finger protein, interacts with the ARD of TRPC6. GST‐pulldown and co‐immunoprecipitation assays showed that RNF24 interacts with all TRPCs. Labeling of the cell surface proteins by biotinylation showed that RNF24 significantly reduced the presence of TRPC at plasma membrane under basal conditions. Also, we observed that the glycosylation pattern of TRPC6 was affected by the expression of RNF24, decreasing the amount of mature glycoprotein and increasing the amount of immature glycoprotein. However, RNF24 did not alter the carbachol‐induced TRPC3 or TRPC6 activation. On the other hand, constitutive calcium entry of TRPC3 was significantly reduced when co‐expressed with RNF24, consistent with a lower basal expression at the cell surface. Using an immunofluorescence approach, we showed that RNF24 is a membrane protein that co‐localizes with mannosidase II, which is an enzyme of the Golgi cisternae. In conclusion, these results indicate that RNF24 interacts with TRPCs at the level of the Golgi cisternae and may be involved in the intracellular trafficking.