TRP channel–associated factors are a novel protein family that regulates TRPM8 trafficking and activity

• DOI: 10.1083/jcb.201402076
• Publication Date: 2015-01-05T17:52:00Z
• Authors: Dimitra Gkika; Loic Lemonnier; George Shapovalov; Dmitri Gordienko; Céline Poux; Michela Bernardini; Alexandre Bokhobza; Gabriel Bidaux; Cindy Degerny; Kathye Verreman; Basma Guarmit; Mohamed Benahmed; Yvan de Launoit; Rene J.M. Bindels; Alessandra Fiorio...; Natalia Prevarskaya
• Keywords: MESH: Signal Transduction; Male; 0301 basic medicine; maximum likelihood method; [SDV]Life Sciences [q-bio]; MESH: TRPM Cation Channels/genetics; TRP channel-associated factor; MESH: Amino Acid Sequence; [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC]; TRP; Abbreviations used in this paper: EAPA2; Membrane Potentials; Cell Movement; MESH: Animals; time-domain fluorescence lifetime imaging microscopy; MESH: Cell Movement; Research Articles; MESH: Middle Aged; MESH: Kinetics; TCAF; LNCaP; Prostate; MESH: Prostatic Neoplasms/pathology; Middle Aged; Radboudumc 11: Renal disorders RIMLS: Radboud Institute for Molecular Life Sciences; 3. Good health; qPCR; Protein Transport; MESH: HEK293 Cells; MESH: Prostate/metabolism*; time-correlated single photon counting; Ion Channel Gating; Protein Binding; MESH: Protein Transport; MESH: Cell Line, Tumor; EAPA2; MESH: RNA Interference; Molecular Sequence Data; human embryonic kidney; [SDV.CAN]Life Sciences [q-bio]/Cancer; Adenocarcinoma; MESH: Adenocarcinoma/pathology; Experimental Autoimmune Prostatitis Antigen 2; MESH: Membrane Proteins/genetics; 03 medical and health sciences; MESH: Adenocarcinoma/genetics; MESH: Mice, Inbred C57BL; Cell Line, Tumor; MESH: Protein Binding; MESH: Membrane Potentials; MESH: Membrane Proteins/metabolism*; Animals; Humans; Neoplasm Invasiveness; quantitative real-time PCR; Amino Acid Sequence; MESH: TRPM Cation Channels/metabolism*; MESH: Prostatic Neoplasms/genetics; HEK; MESH: Humans; MESH: Molecular Sequence Data; [SDV.GEN.GPO]Life Sciences [q-bio]/Genetics/Populations and Evolution [q-bio.PE]; MESH: Prostate/pathology; TCSPC; MESH: Transfection; MESH: Adenocarcinoma/metabolism*; Membrane Proteins; Prostatic Neoplasms; MESH: Neoplasm Invasiveness; MESH: Prostatic Neoplasms/metabolism*; MESH: Ion Channel Gating; ML; MESH: Male; Mice, Inbred C57BL; Kinetics; HEK293 Cells; lymph node carcinoma of prostate; Förster resonance energy transfer; transient receptor potential; FRET; [SDV.GEN.GPO] Life Sciences [q-bio]/Genetics/Populations and Evolution [q-bio.PE]; TD FLIM

TRPM8 is a cold sensor that is highly expressed in the prostate as well as in other non-temperature-sensing organs, and is regulated by downstream receptor–activated signaling pathways. However, little is known about the intracellular proteins necessary for channel function. Here, we identify two previously unknown proteins, which we have named “TRP channel–associated factors” (TCAFs), as new TRPM8 partner proteins, and we demonstrate that they are necessary for channel function. TCAF1 and TCAF2 both bind to the TRPM8 channel and promote its trafficking to the cell surface. However, they exert opposing effects on TRPM8 gating properties. Functional interaction of TCAF1/TRPM8 also leads to a reduction in both the speed and directionality of migration of prostate cancer cells, which is consistent with an observed loss of expression of TCAF1 in metastatic human specimens, whereas TCAF2 promotes migration. The identification of TCAFs introduces a novel mechanism for modulation of TRPM8 channel activity.