SNF8, a member of the ESCRT-II complex, interacts with TRPC6 and enhances its channel activity

BMC Cell Biology

Research Article

Dequan Tian¹  Anna Greka¹  Martin R Pollak²  Johannes Schlöndorff²  Sneha Krishna²  Robert Carrasquillo² 

[1] Department of Medicine, Nephrology Division, Massachusetts General Hospital, 149 13th Street, Room 8.102, 02114, Boston, MA, USA;Harvard Medical School, 02115, Boston, MA, USA;Division of Nephrology, Beth Israel Deaconess Medical Center, Research North 304B, 99 Brookline Ave, 02215, Boston, MA, USA;Harvard Medical School, 02115, Boston, MA, USA;

关键词: Transient receptor potential;    Calcium channel;    Protein-protein interaction;    Calcineurin-NFAT signaling;   

DOI  :  10.1186/1471-2121-13-33

 received in 2012-07-10, accepted in 2012-10-23,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundTransient receptor potential canonical (TRPC) channels are non-selective cation channels involved in receptor-mediated calcium signaling in diverse cells and tissues. The canonical transient receptor potential 6 (TRPC6) has been implicated in several pathological processes, including focal segmental glomerulosclerosis (FSGS), cardiac hypertrophy, and pulmonary hypertension. The two large cytoplasmic segments of the cation channel play a critical role in the proper regulation of channel activity, and are involved in several protein-protein interactions.ResultsHere we report that SNF8, a component of the endosomal sorting complex for transport-II (ESCRT-II) complex, interacts with TRPC6. The interaction was initially observed in a yeast two-hybrid screen using the amino-terminal cytoplasmic domain of TRPC6 as bait, and confirmed by co-immunoprecipitation from eukaryotic cell extracts. The amino-terminal 107 amino acids are necessary and sufficient for the interaction. Overexpression of SNF8 enhances both wild-type and gain-of-function mutant TRPC6-mediated whole-cell currents in HEK293T cells. Furthermore, activation of NFAT-mediated transcription by gain-of-function mutants is enhanced by overexpression of SNF8, and partially inhibited by RNAi mediated knockdown of SNF8. Although the ESCRT-II complex functions in the endocytosis and lysosomal degradation of transmembrane proteins, SNF8 overexpression does not alter the amount of TRPC6 present on the cell surface.ConclusionSNF8 is novel binding partner of TRPC6, binding to the amino-terminal cytoplasmic domain of the channel. Modulating SNF8 expression levels alters the TRPC6 channel current and can modulate activation of NFAT-mediated transcription downstream of gain-of-function mutant TRPC6. Taken together, these results identify SNF8 as a novel regulator of TRPC6.

【 授权许可】

Unknown   
© Carrasquillo et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.