EEPD1 Is a Novel LXR Target Gene in Macrophages Which Regulates ABCA1 Abundance and Cholesterol Efflux

The sterol-responsive nuclear receptors, liver X receptors α (LXRα, ) and β (LXRβ, ), are key determinants of cellular cholesterol homeostasis. LXRs are activated under conditions of high cellular sterol load and induce expression of the cholesterol efflux transporters and to promote efflux of exces...

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Vydáno v:Arteriosclerosis, thrombosis, and vascular biology Ročník 37; číslo 3; s. 423 - 432
Hlavní autoři: Nelson, Jessica Kristine, Koenis, Duco Steven, Scheij, Saskia, Cook, Emma Clare Laura, Moeton, Martina, Santos, Ana, Lobaccaro, Jean-Marc Adolphe, Baron, Silvere, Zelcer, Noam
Médium: Journal Article
Jazyk:angličtina
Vydáno: United States 01.03.2017
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ISSN:1524-4636
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Abstract The sterol-responsive nuclear receptors, liver X receptors α (LXRα, ) and β (LXRβ, ), are key determinants of cellular cholesterol homeostasis. LXRs are activated under conditions of high cellular sterol load and induce expression of the cholesterol efflux transporters and to promote efflux of excess cellular cholesterol. However, the full set of genes that contribute to LXR-stimulated cholesterol efflux is unknown, and their identification is the objective of this study. We systematically compared the global transcriptional response of macrophages to distinct classes of LXR ligands. This allowed us to identify both common and ligand-specific transcriptional responses in macrophages. Among these, we identified endonuclease-exonuclease-phosphatase family domain containing 1 ( ) as a direct transcriptional target of LXRs in human and murine macrophages. EEPD1 specifically localizes to the plasma membrane owing to the presence of a myristoylation site in its N terminus. Accordingly, the first 10 amino acids of EEPD1 are sufficient to confer plasma membrane localization in the context of a chimeric protein with GFP. Functionally, we report that silencing expression of blunts maximal LXR-stimulated Apo AI-dependent efflux and demonstrate that this is the result of reduced abundance of ABCA1 protein in human and murine macrophages. In this study, we identify as a novel LXR-regulated gene in macrophages and propose that it promotes cellular cholesterol efflux by controlling cellular levels and activity of ABCA1.
AbstractList The sterol-responsive nuclear receptors, liver X receptors α (LXRα, ) and β (LXRβ, ), are key determinants of cellular cholesterol homeostasis. LXRs are activated under conditions of high cellular sterol load and induce expression of the cholesterol efflux transporters and to promote efflux of excess cellular cholesterol. However, the full set of genes that contribute to LXR-stimulated cholesterol efflux is unknown, and their identification is the objective of this study. We systematically compared the global transcriptional response of macrophages to distinct classes of LXR ligands. This allowed us to identify both common and ligand-specific transcriptional responses in macrophages. Among these, we identified endonuclease-exonuclease-phosphatase family domain containing 1 ( ) as a direct transcriptional target of LXRs in human and murine macrophages. EEPD1 specifically localizes to the plasma membrane owing to the presence of a myristoylation site in its N terminus. Accordingly, the first 10 amino acids of EEPD1 are sufficient to confer plasma membrane localization in the context of a chimeric protein with GFP. Functionally, we report that silencing expression of blunts maximal LXR-stimulated Apo AI-dependent efflux and demonstrate that this is the result of reduced abundance of ABCA1 protein in human and murine macrophages. In this study, we identify as a novel LXR-regulated gene in macrophages and propose that it promotes cellular cholesterol efflux by controlling cellular levels and activity of ABCA1.
OBJECTIVEThe sterol-responsive nuclear receptors, liver X receptors α (LXRα, NR1H3) and β (LXRβ, NR1H2), are key determinants of cellular cholesterol homeostasis. LXRs are activated under conditions of high cellular sterol load and induce expression of the cholesterol efflux transporters ABCA1 and ABCG1 to promote efflux of excess cellular cholesterol. However, the full set of genes that contribute to LXR-stimulated cholesterol efflux is unknown, and their identification is the objective of this study.APPROACH AND RESULTSWe systematically compared the global transcriptional response of macrophages to distinct classes of LXR ligands. This allowed us to identify both common and ligand-specific transcriptional responses in macrophages. Among these, we identified endonuclease-exonuclease-phosphatase family domain containing 1 (EEPD1/KIAA1706) as a direct transcriptional target of LXRs in human and murine macrophages. EEPD1 specifically localizes to the plasma membrane owing to the presence of a myristoylation site in its N terminus. Accordingly, the first 10 amino acids of EEPD1 are sufficient to confer plasma membrane localization in the context of a chimeric protein with GFP. Functionally, we report that silencing expression of EEPD1 blunts maximal LXR-stimulated Apo AI-dependent efflux and demonstrate that this is the result of reduced abundance of ABCA1 protein in human and murine macrophages.CONCLUSIONSIn this study, we identify EEPD1 as a novel LXR-regulated gene in macrophages and propose that it promotes cellular cholesterol efflux by controlling cellular levels and activity of ABCA1.
Author Cook, Emma Clare Laura
Lobaccaro, Jean-Marc Adolphe
Nelson, Jessica Kristine
Scheij, Saskia
Baron, Silvere
Zelcer, Noam
Koenis, Duco Steven
Santos, Ana
Moeton, Martina
Author_xml – sequence: 1
  givenname: Jessica Kristine
  surname: Nelson
  fullname: Nelson, Jessica Kristine
  organization: From the Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands (J.K.N., D.S.K., S.S., E.C.L.C., M.M., A.S., N.Z.); and Université Clermont Auvergne, CNRS, Inserm, GReD, Clermont-Ferrand, France (J.-M.A.L., S.B.)
– sequence: 2
  givenname: Duco Steven
  surname: Koenis
  fullname: Koenis, Duco Steven
  organization: From the Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands (J.K.N., D.S.K., S.S., E.C.L.C., M.M., A.S., N.Z.); and Université Clermont Auvergne, CNRS, Inserm, GReD, Clermont-Ferrand, France (J.-M.A.L., S.B.)
– sequence: 3
  givenname: Saskia
  surname: Scheij
  fullname: Scheij, Saskia
  organization: From the Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands (J.K.N., D.S.K., S.S., E.C.L.C., M.M., A.S., N.Z.); and Université Clermont Auvergne, CNRS, Inserm, GReD, Clermont-Ferrand, France (J.-M.A.L., S.B.)
– sequence: 4
  givenname: Emma Clare Laura
  surname: Cook
  fullname: Cook, Emma Clare Laura
  organization: From the Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands (J.K.N., D.S.K., S.S., E.C.L.C., M.M., A.S., N.Z.); and Université Clermont Auvergne, CNRS, Inserm, GReD, Clermont-Ferrand, France (J.-M.A.L., S.B.)
– sequence: 5
  givenname: Martina
  surname: Moeton
  fullname: Moeton, Martina
  organization: From the Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands (J.K.N., D.S.K., S.S., E.C.L.C., M.M., A.S., N.Z.); and Université Clermont Auvergne, CNRS, Inserm, GReD, Clermont-Ferrand, France (J.-M.A.L., S.B.)
– sequence: 6
  givenname: Ana
  surname: Santos
  fullname: Santos, Ana
  organization: From the Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands (J.K.N., D.S.K., S.S., E.C.L.C., M.M., A.S., N.Z.); and Université Clermont Auvergne, CNRS, Inserm, GReD, Clermont-Ferrand, France (J.-M.A.L., S.B.)
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  givenname: Jean-Marc Adolphe
  surname: Lobaccaro
  fullname: Lobaccaro, Jean-Marc Adolphe
  organization: From the Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands (J.K.N., D.S.K., S.S., E.C.L.C., M.M., A.S., N.Z.); and Université Clermont Auvergne, CNRS, Inserm, GReD, Clermont-Ferrand, France (J.-M.A.L., S.B.)
– sequence: 8
  givenname: Silvere
  surname: Baron
  fullname: Baron, Silvere
  organization: From the Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands (J.K.N., D.S.K., S.S., E.C.L.C., M.M., A.S., N.Z.); and Université Clermont Auvergne, CNRS, Inserm, GReD, Clermont-Ferrand, France (J.-M.A.L., S.B.)
– sequence: 9
  givenname: Noam
  surname: Zelcer
  fullname: Zelcer, Noam
  email: n.zelcer@amc.uva.nl
  organization: From the Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, The Netherlands (J.K.N., D.S.K., S.S., E.C.L.C., M.M., A.S., N.Z.); and Université Clermont Auvergne, CNRS, Inserm, GReD, Clermont-Ferrand, France (J.-M.A.L., S.B.). n.zelcer@amc.uva.nl
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Keywords ABCA1
nuclear receptors
macrophages
cholesterol efflux
cholesterol metabolism
LXR
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Snippet The sterol-responsive nuclear receptors, liver X receptors α (LXRα, ) and β (LXRβ, ), are key determinants of cellular cholesterol homeostasis. LXRs are...
OBJECTIVEThe sterol-responsive nuclear receptors, liver X receptors α (LXRα, NR1H3) and β (LXRβ, NR1H2), are key determinants of cellular cholesterol...
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SubjectTerms Animals
Apolipoprotein A-I - metabolism
ATP Binding Cassette Transporter 1 - genetics
ATP Binding Cassette Transporter 1 - metabolism
Biological Transport
Cell Membrane - drug effects
Cell Membrane - enzymology
Chlorocebus aethiops
Cholesterol - metabolism
COS Cells
Endodeoxyribonucleases - genetics
Endodeoxyribonucleases - metabolism
Gene Expression Profiling - methods
Gene Expression Regulation, Enzymologic
HeLa Cells
Hep G2 Cells
Humans
Ligands
Liver X Receptors - agonists
Liver X Receptors - deficiency
Liver X Receptors - genetics
Liver X Receptors - metabolism
Macrophages - drug effects
Macrophages - enzymology
Male
Mice
Mice, Inbred C57BL
Mice, Knockout
RAW 264.7 Cells
RNA Interference
Transcriptome
Transfection
Title EEPD1 Is a Novel LXR Target Gene in Macrophages Which Regulates ABCA1 Abundance and Cholesterol Efflux
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