title: The deubiquitinating enzyme USP5 modulates neuropathic and inflammatory pain by enhancing Cav3.2 channel activity.
abstract: T-type calcium channels are essential contributors to the transmission of nociceptive signals in the primary afferent pain pathway. Here, we show that T-type calcium channels are ubiquitinated by WWP1, a plasma-membrane-associated ubiquitin ligase that binds to the intracellular domain III-IV linker region of the Cav3.2 T-type channel and modifies specific lysine residues in this region. A proteomic screen identified the deubiquitinating enzyme USP5 as a Cav3.2 III-IV linker interacting partner. Knockdown of USP5 via shRNA increases Cav3.2 ubiquitination, decreases Cav3.2 protein levels, and reduces Cav3.2 whole-cell currents. In vivo knockdown of USP5 or uncoupling USP5 from native Cav3.2 channels via intrathecal delivery of Tat peptides mediates analgesia in both inflammatory and neuropathic mouse models of mechanical hypersensitivity. Altogether, our experiments reveal a cell signaling pathway that regulates T-type channel activity and their role in nociceptive signaling.
authors.affiliation: Department of Physiology and Pharmacology, Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Agustin
authors.initials: A
authors.lastname: García-Caballero
authors.affiliation: Department of Physiology and Pharmacology, Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Vinicius M
authors.initials: VM
authors.lastname: Gadotti
authors.affiliation: Department of Physiology and Pharmacology, Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Patrick
authors.initials: P
authors.lastname: Stemkowski
authors.affiliation: Department of Physiology and Pharmacology, Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Norbert
authors.initials: N
authors.lastname: Weiss
authors.affiliation: Department of Physiology and Pharmacology, Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Ivana A
authors.initials: IA
authors.lastname: Souza
authors.affiliation: Department of Physiology and Pharmacology, Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Victoria
authors.initials: V
authors.lastname: Hodgkinson
authors.affiliation: Department of Physiology and Pharmacology, Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Chris
authors.initials: C
authors.lastname: Bladen
authors.affiliation: Department of Physiology and Pharmacology, Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Lina
authors.initials: L
authors.lastname: Chen
authors.affiliation: Department of Physiology and Pharmacology, Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Jawed
authors.initials: J
authors.lastname: Hamid
authors.affiliation: Laboratories of Excellence in Ion Channel Science and Therapeutics, Institut de Génomique Fonctionnelle, CNRS UMR5203, INSERM U661, IFR3 Universités Montpellier I&II, Montpellier, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Pizzoccaro
authors.affiliation: Laboratories of Excellence in Ion Channel Science and Therapeutics, Institut de Génomique Fonctionnelle, CNRS UMR5203, INSERM U661, IFR3 Universités Montpellier I&II, Montpellier, France.
authors.forename: Mickael
authors.initials: M
authors.lastname: Deage
authors.affiliation: Laboratories of Excellence in Ion Channel Science and Therapeutics, Institut de Génomique Fonctionnelle, CNRS UMR5203, INSERM U661, IFR3 Universités Montpellier I&II, Montpellier, France.
authors.forename: Amaury
authors.initials: A
authors.lastname: François
authors.affiliation: Laboratories of Excellence in Ion Channel Science and Therapeutics, Institut de Génomique Fonctionnelle, CNRS UMR5203, INSERM U661, IFR3 Universités Montpellier I&II, Montpellier, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Bourinet
authors.affiliation: Department of Physiology and Pharmacology, Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada. Electronic address: Zamponi@ucalgary.ca.
authors.forename: Gerald W
authors.initials: GW
authors.lastname: Zamponi
mesh_terms: D000818:Animals; D020747:Calcium Channels, T-Type / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D004195:Disease Models, Animal; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D005620:Freund's Adjuvant / Q000633:toxicity; D006801:Humans; D006930:Hyperalgesia / Q000175:diagnosis / Q000503:physiopathology; D066298:In Vitro Techniques; D007249:Inflammation / Q000139:chemically induced / Q000503:physiopathology*; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000235:genetics; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009437:Neuralgia / Q000188:drug therapy / Q000201:enzymology*; D017288:Pain Threshold / Q000187:drug effects / Q000502:physiology; D010455:Peptides / Q000627:therapeutic use; D011984:Sensory Receptor Cells / Q000187:drug effects / Q000502:physiology; D013116:Spinal Cord / Q000166:cytology; D014162:Transfection; D054875:Ubiquitination / Q000235:genetics / Q000502:physiology
chemical_list: C504883:Cacna1h protein, mouse; D020747:Calcium Channels, T-Type; D009419:Nerve Tissue Proteins; D010455:Peptides; D005620:Freund's Adjuvant; D010450:Endopeptidases; C091939:ubiquitin isopeptidase