Coordinated regulation of the orosomucoid-like gene family expression controls de novo ceramide synthesis in mammalian cells
Mostra el registre complet Registre parcial de l'ítem
- dc.contributor.author Kiefer, Kerstin, 1986-
- dc.contributor.author Carreras Sureda, Amado, 1986-
- dc.contributor.author García López, Roberto, 1991-
- dc.contributor.author Rubio Moscardó, Fanny
- dc.contributor.author Casas, Josefina
- dc.contributor.author Fabriàs, Gemma
- dc.contributor.author Vicente García, Rubén, 1978-
- dc.date.accessioned 2024-02-19T10:07:55Z
- dc.date.available 2024-02-19T10:07:55Z
- dc.date.issued 2015
- dc.description.abstract The orosomucoid-like (ORMDL) protein family is involved in the regulation of de novo sphingolipid synthesis, calcium homeostasis, and unfolded protein response. Single nucleotide polymorphisms (SNPs) that increase ORMDL3 expression have been associated with various immune/inflammatory diseases, although the pathophysiological mechanisms underlying this association are poorly understood. ORMDL proteins are claimed to be inhibitors of the serine palmitoyltransferase (SPT). However, it is not clear whether individual ORMDL expression levels have an impact on ceramide synthesis. The present study addressed the interaction with and regulation of SPT activity by ORMDLs to clarify their pathophysiological relevance. We have measured ceramide production in HEK293 cells incubated with palmitate as a direct substrate for SPT reaction. Our results showed that a coordinated overexpression of the three isoforms inhibits the enzyme completely, whereas individual ORMDLs are not as effective. Immunoprecipitation and fluorescence resonance energy transfer (FRET) studies showed that mammalian ORMDLs form oligomeric complexes that change conformation depending on cellular sphingolipid levels. Finally, using macrophages as a model, we demonstrate that mammalian cells modify ORMDL genes expression levels coordinately to regulate the de novo ceramide synthesis pathway. In conclusion, we have shown a physiological modulation of SPT activity by general ORMDL expression level regulation. Moreover, because single ORMDL3 protein alteration produces an incomplete inhibition of SPT activity, this work argues against the idea that ORMDL3 pathophysiology could be explained by a simple on/off mechanism on SPT activity.
- dc.description.sponsorship This work was supported by Spanish Ministry of Economy and Competitiveness Grants SAF2010-16725 and SAF2011-22444, Fondo de Investigación Sanitaria Red HERACLES Grant RD12/0042/0014, Fondos Europeos de Desarrollo Regional, Generalitat de Catalunya Grant SGR2014, and Fundació la Marató de TV3 Grant 20134030.
- dc.format.mimetype application/pdf
- dc.identifier.citation Kiefer K, Carreras-Sureda A, García-López R, Rubio-Moscardó F, Casas J, Fabriàs G, et al. Coordinated regulation of the orosomucoid-like gene family expression controls de novo ceramide synthesis in mammalian cells. Journal of Biological Chemistry. 2015 Jan 30;290(5):2822-30. DOI: 10.1074/jbc.M114.595116
- dc.identifier.doi http://dx.doi.org/10.1074/jbc.M114.595116
- dc.identifier.issn 0021-9258
- dc.identifier.uri http://hdl.handle.net/10230/59138
- dc.language.iso eng
- dc.publisher Elsevier
- dc.relation.ispartof Journal of Biological Chemistry. 2015 Jan 30;290(5):2822-30
- dc.relation.projectID info:eu-repo/grantAgreement/ES/3PN/SAF2010-16725
- dc.relation.projectID info:eu-repo/grantAgreement/ES/3PN/SAF2011-22444
- dc.rights © 2015 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology. This is an open access article distributed under the terms of the Creative Commons CC-BY license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
- dc.rights.accessRights info:eu-repo/semantics/openAccess
- dc.rights.uri http://creativecommons.org/licenses/by/4.0/
- dc.subject.keyword Ceramide
- dc.subject.keyword Enzyme inhibitor
- dc.subject.keyword Macrophage
- dc.subject.keyword Serine palmitoyltransferase
- dc.subject.keyword Sphingolipid
- dc.title Coordinated regulation of the orosomucoid-like gene family expression controls de novo ceramide synthesis in mammalian cells
- dc.type info:eu-repo/semantics/article
- dc.type.version info:eu-repo/semantics/publishedVersion