title: Interaction between the Linker, Pre-S1, and TRP Domains Determines Folding, Assembly, and Trafficking of TRPV Channels.
abstract: Functional transient receptor potential (TRP) channels result from the assembly of four subunits. Here, we show an interaction between the pre-S1, TRP, and the ankyrin repeat domain (ARD)-S1 linker domains of TRPV1 and TRPV4 that is essential for proper channel assembly. Neutralization of TRPV4 pre-S1 K462 resulted in protein retention in the ER, defective glycosylation and trafficking, and unresponsiveness to TRPV4-activating stimuli. Similar results were obtained with the equivalent mutation in TRPV1 pre-S1. Molecular dynamics simulations revealed that TRPV4-K462 generated an alternating hydrogen network with E745 (TRP box) and D425 (pre-S1 linker), and that K462Q mutation affected subunit folding. Consistently, single TRPV4-E745A or TRPV4-D425A mutations moderately affected TRPV4 biogenesis while double TRPV4-D425A/E745A mutation resumed the TRPV4-K462Q phenotype. Thus, the interaction between pre-S1, TRP, and linker domains is mandatory to generate a structural conformation that allows the contacts between adjacent subunits to promote correct assembly and trafficking to the plasma membrane.
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona 08003, Spain.
authors.forename: Anna
authors.initials: A
authors.lastname: Garcia-Elias
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona 08003, Spain.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Berna-Erro
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona 08003, Spain.
authors.forename: Fanny
authors.initials: F
authors.lastname: Rubio-Moscardo
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona 08003, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Pardo-Pastor
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona 08003, Spain.
authors.forename: Sanela
authors.initials: S
authors.lastname: Mrkonjić
authors.affiliation: Universidad Andrés Bello, Center for Bioinformatics and Integrative Biology, Facultad de Ciencias Biológicas, Av. República 239, Santiago 8320000, Chile; Centro Interdisciplinario de Neurociencia de Valparaíso, Facultad de Ciencias, Universidad de Valparaíso, Valparaíso 2366103, Chile.
authors.forename: Romina V
authors.initials: RV
authors.lastname: Sepúlveda
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona 08003, Spain.
authors.forename: Rubén
authors.initials: R
authors.lastname: Vicente
authors.affiliation: Universidad Andrés Bello, Center for Bioinformatics and Integrative Biology, Facultad de Ciencias Biológicas, Av. República 239, Santiago 8320000, Chile; Centro Interdisciplinario de Neurociencia de Valparaíso, Facultad de Ciencias, Universidad de Valparaíso, Valparaíso 2366103, Chile.
authors.forename: Fernando
authors.initials: F
authors.lastname: González-Nilo
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona 08003, Spain. Electronic address: miguel.valverde@upf.edu.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Valverde
mesh_terms: D000595:Amino Acid Sequence; D015870:Gene Expression; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D006860:Hydrogen Bonding; D008564:Membrane Potentials / Q000502:physiology; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D009154:Mutation; D017510:Protein Folding; D054730:Protein Interaction Domains and Motifs; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D016415:Sequence Alignment; D050916:TRPV Cation Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D021122:Protein Subunits; D050916:TRPV Cation Channels; C494482:TRPV1 protein, human; C416986:TRPV4 protein, human