title: Regulation of TRPC6 channel activity by tyrosine phosphorylation.
abstract: Various hormonal stimuli and growth factors activate the mammalian canonical transient receptor potential (TRPC) channel through phospholipase C (PLC) activation. However, the precise mechanism of the regulation of TRPC channel activity remains unknown. Here, we provide the first evidence that direct tyrosine phosphorylation by Src family protein-tyrosine kinases (PTKs) is a novel mechanism for modulating TRPC6 channel activity. We found that TRPC6 is tyrosine-phosphorylated in COS-7 cells when coexpressed with Fyn, a member of the Src family PTKs. We also found that Fyn interacts with TRPC6 and that the interaction is mediated by the SH2 domain of Fyn and the N-terminal region of TRPC6 in a phosphorylation-independent manner. In addition, we demonstrated the physical association of TRPC6 with Fyn in the mammalian brain. Moreover, we showed that stimulation of the epidermal growth factor receptor induced rapid tyrosine phosphorylation of TRPC6 in COS-7 cells. This epidermal growth factor-induced tyrosine phosphorylation of TRPC6 was significantly blocked by PP2, a specific inhibitor of Src family PTKs, and by a dominant negative form of Fyn, suggesting that the direct phosphorylation of TRPC6 by Src family PTKs could be caused by physiological stimulation. Furthermore, using single channel recording, we showed that Fyn modulates TRPC6 channel activity via tyrosine phosphorylation. Thus, our findings demonstrated that tyrosine phosphorylation by Src family PTKs is a novel regulatory mechanism of TRPC6 channel activity.
authors.affiliation: Laboratory for Developmental Neurobiology, RIKEN Brain Science Institute (BSI), 2-1 Hirosawa, Wako City, Saitama 351-0198, Japan. chihiro@brain.riken.go.jp
authors.forename: Chihiro
authors.initials: C
authors.lastname: Hisatsune
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Kuroda
authors.forename: Kyoko
authors.initials: K
authors.lastname: Nakamura
authors.forename: Takafumi
authors.initials: T
authors.lastname: Inoue
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nakamura
authors.forename: Takayuki
authors.initials: T
authors.lastname: Michikawa
authors.forename: Akihiro
authors.initials: A
authors.lastname: Mizutani
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Mikoshiba
mesh_terms: D000818:Animals; D001921:Brain; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D066246:ErbB Receptors / Q000378:metabolism; D051379:Mice; D008861:Microsomes / Q000737:chemistry; D010766:Phosphorylation; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051076:Proto-Oncogene Proteins c-fyn; D051381:Rats; D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D015220:Calcium Channels; D011518:Proto-Oncogene Proteins; D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; C492146:Trpc6 protein, mouse; C492145:Trpc6 protein, rat; D014443:Tyrosine; D066246:ErbB Receptors; C494960:Fyn protein, mouse; C494961:Fyn protein, rat; D051076:Proto-Oncogene Proteins c-fyn; D019061:src-Family Kinases; D002118:Calcium