TY - JOUR
T1 - TRPM4 is a novel component of the adhesome required for focal adhesion disassembly, migration and contractility
AU - Cáceres, Mónica
AU - Ortiz, Liliana
AU - Recabarren, Tatiana
AU - Romero, Anibal
AU - Colombo, Alicia
AU - Leiva-Salcedo, Elías
AU - Varela, Diego
AU - Rivas, José
AU - Silva, Ian
AU - Morales, Diego
AU - Campusano, Camilo
AU - Almarza, Oscar
AU - Simon, Felipe
AU - Toledo, Hector
AU - Park, Kang Sik
AU - Trimmer, James S.
AU - Cerda, Oscar
N1 - Funding Information:
We are thankful to Mr. Nicanor Villarroel, Mrs. Irma Orellana, Mr. Miladio Ruz and members of the Fish Facility for technical support. We thank Dr. Pierre Launay for kindly providing the pcDNA4TO-FLAG-hTRPM4 plasmid. We are also grateful to Dr. Christopher Turner for providing the EGFP-Paxillin plasmid, and Dr. Alan Hall for the pRK5 Rac1 L61 plasmid (plasmid 15904 via Addgene, Cambridge, MA, USA). Mass spectrometry was performed at the University of California Davis Proteomics Facility. We thank Dr. Marcelo Catalán for his help in Ca imaging experiments, and Drs. Dave Speca and Aldo Solari for constructive discussions related to the manuscript. We also thank to Drs. Remigio López and Benjamín Suárez for their support during the initial stages of this project. This research was funded by Fondecyt 11121239 to OC, Fondecyt 11140064 to MC, and NIH R01 NS042225 to JST. A doctoral Fellowship from Conicyt supports AR Fondecyt 1120240 funded DV Fondecyt 1121078 and Millennium Institute on Immunology and Immunotherapy P09-016-F supported FS. Fondecyt 1120126 funded HT. 2+
Publisher Copyright:
© 2015, Public Library of Science. All rights reserved. This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.
PY - 2015/6/25
Y1 - 2015/6/25
N2 - Cellular migration and contractility are fundamental processes that are regulated by a variety of concerted mechanisms such as cytoskeleton rearrangements, focal adhesion turnover, and Ca2+ oscillations. TRPM4 is a Ca2+-activated non-selective cationic channel (Ca2+-NSCC) that conducts monovalent but not divalent cations. Here, we used a mass spectrometry-based proteomics approach to identify putative TRPM4-associated proteins. Interestingly, the largest group of these proteins has actin cytoskeleton-related functions, and among these nine are specifically annotated as focal adhesion-related proteins. Consistent with these results, we found that TRPM4 localizes to focal adhesions in cells from different cellular lineages. We show that suppression of TRPM4 in MEFs impacts turnover of focal adhesions, serum-induced Ca2+ influx, focal adhesion kinase (FAK) and Rac activities, and results in reduced cellular spreading, migration and contractile behavior. Finally, we demonstrate that the inhibition of TRPM4 activity alters cellular contractility in vivo, affecting cutaneous wound healing. Together, these findings provide the first evidence, to our knowledge, for a TRP channel specifically localized to focal adhesions, where it performs a central role in modulating cellular migration and contractility.
AB - Cellular migration and contractility are fundamental processes that are regulated by a variety of concerted mechanisms such as cytoskeleton rearrangements, focal adhesion turnover, and Ca2+ oscillations. TRPM4 is a Ca2+-activated non-selective cationic channel (Ca2+-NSCC) that conducts monovalent but not divalent cations. Here, we used a mass spectrometry-based proteomics approach to identify putative TRPM4-associated proteins. Interestingly, the largest group of these proteins has actin cytoskeleton-related functions, and among these nine are specifically annotated as focal adhesion-related proteins. Consistent with these results, we found that TRPM4 localizes to focal adhesions in cells from different cellular lineages. We show that suppression of TRPM4 in MEFs impacts turnover of focal adhesions, serum-induced Ca2+ influx, focal adhesion kinase (FAK) and Rac activities, and results in reduced cellular spreading, migration and contractile behavior. Finally, we demonstrate that the inhibition of TRPM4 activity alters cellular contractility in vivo, affecting cutaneous wound healing. Together, these findings provide the first evidence, to our knowledge, for a TRP channel specifically localized to focal adhesions, where it performs a central role in modulating cellular migration and contractility.
UR - http://www.scopus.com/inward/record.url?scp=84938630783&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0130540
DO - 10.1371/journal.pone.0130540
M3 - Review article
C2 - 26110647
AN - SCOPUS:84938630783
SN - 1932-6203
VL - 10
JO - PLoS ONE
JF - PLoS ONE
IS - 6
M1 - e0130540
ER -