title: Inhibition of ubiquitin proteasome system rescues the defective sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA1) protein causing Chianina cattle pseudomyotonia.
abstract: A missense mutation in ATP2A1 gene, encoding sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA1) protein, causes Chianina cattle congenital pseudomyotonia, an exercise-induced impairment of muscle relaxation. Skeletal muscles of affected cattle are characterized by a selective reduction of SERCA1 in sarcoplasmic reticulum membranes. In this study, we provide evidence that the ubiquitin proteasome system is involved in the reduced density of mutated SERCA1. The treatment with MG132, an inhibitor of ubiquitin proteasome system, rescues the expression level and membrane localization of the SERCA1 mutant in a heterologous cellular model. Cells co-transfected with the Ca(2+)-sensitive probe aequorin show that the rescued SERCA1 mutant exhibits the same ability of wild type to maintain Ca(2+) homeostasis within cells. These data have been confirmed by those obtained ex vivo on adult skeletal muscle fibers from a biopsy from a pseudomyotonia-affected subject. Our data show that the mutation generates a protein most likely corrupted in proper folding but not in catalytic activity. Rescue of mutated SERCA1 to sarcoplasmic reticulum membrane can re-establish resting cytosolic Ca(2+) concentration and prevent the appearance of pathological signs of cattle pseudomyotonia.
authors.affiliation: From the Departments of Biomedical Sciences and.
authors.forename: Elisa
authors.initials: E
authors.lastname: Bianchini
authors.affiliation: the Departments of Animal Medicine Production and Health and.
authors.forename: Stefania
authors.initials: S
authors.lastname: Testoni
authors.affiliation: the Department of Veterinary Medical Sciences, University of Bologna, 40064 Bologna, Italy.
authors.forename: Arcangelo
authors.initials: A
authors.lastname: Gentile
authors.affiliation: Biology, University of Padova, 35131 Padova, Italy.
authors.forename: Tito
authors.initials: T
authors.lastname: Calì
authors.affiliation: Biology, University of Padova, 35131 Padova, Italy.
authors.forename: Denis
authors.initials: D
authors.lastname: Ottolini
authors.affiliation: the Consorzio M.I.A., University of Milano Bicocca, 20900 Monza, Italy.
authors.forename: Antonello
authors.initials: A
authors.lastname: Villa
authors.affiliation: Biology, University of Padova, 35131 Padova, Italy.
authors.forename: Marisa
authors.initials: M
authors.lastname: Brini
authors.affiliation: the Neuroscience Institute, Consiglio Nazionale delle Ricerche Padova, 35131 Padova, Italy, and.
authors.forename: Romeo
authors.initials: R
authors.lastname: Betto
authors.affiliation: Comparative Biomedicine and Food Science, University of Padova,35020 Legnaro (Padova), Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Mascarello
authors.affiliation: the Department of Molecular Biology and Genetics, Centre for Membrane Pumps in Cells and Disease, Aarhus University, 8000 Aarhus, Denmark.
authors.forename: Poul
authors.initials: P
authors.lastname: Nissen
authors.affiliation: From the Departments of Biomedical Sciences and dorianna.sandona@unipd.it.
authors.forename: Dorianna
authors.initials: D
authors.lastname: Sandonà
authors.affiliation: Comparative Biomedicine and Food Science, University of Padova,35020 Legnaro (Padova), Italy, roberta.sacchetto@unipd.it.
authors.forename: Roberta
authors.initials: R
authors.lastname: Sacchetto
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002418:Cattle Diseases / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D006224:Cricetinae; D057809:HEK293 Cells; D006801:Humans; D020386:Isaacs Syndrome / Q000201:enzymology* / Q000235:genetics / Q000473:pathology / Q000662:veterinary*; D007976:Leupeptins / Q000494:pharmacology; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D061988:Proteasome Inhibitors / Q000494:pharmacology; D017510:Protein Folding / Q000187:drug effects; D012519:Sarcoplasmic Reticulum / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000235:genetics / Q000378:metabolism*; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism*
chemical_list: D007976:Leupeptins; D009124:Muscle Proteins; D061988:Proteasome Inhibitors; D025801:Ubiquitin; D046988:Proteasome Endopeptidase Complex; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde; D002118:Calcium
keywords: Brody Disease; Calcium; Calcium ATPase; Cattle Congenital Pseudomyotonia; Proteasome; Sarco(endo)plasmic Reticulum Calcium ATPase (SERCA1); Sarcoplasmic Reticulum (SR); Skeletal Muscle