title: Oxysterol-binding protein (OSBP)-related protein 4 (ORP4) is essential for cell proliferation and survival.
abstract: Oxysterol-binding protein (OSBP) and OSBP-related proteins (ORPs) comprise a large gene family with sterol/lipid transport and regulatory activities. ORP4 (OSBP2) is a closely related paralogue of OSBP, but its function is unknown. Here we show that ORP4 binds similar sterol and lipid ligands as OSBP and other ORPs but is uniquely required for the proliferation and survival of cultured cells. Recombinant ORP4L and a variant without a pleckstrin homology (PH) domain (ORP4S) bind 25-hydroxycholesterol and extract and transfer cholesterol between liposomes. Two conserved histidine residues in the OSBP homology domain ORP4 are essential for binding phosphatidylinositol 4-phosphate but not sterols. The PH domain of ORP4L also binds phosphatidylinositol 4-phosphate in the Golgi apparatus. However, in the context of ORP4L, the PH domain is required for normal organization of the vimentin network. Unlike OSBP, RNAi silencing of all ORP4 variants (including a partial PH domain truncation termed ORP4M) in HEK293 and HeLa cells resulted in growth arrest but not cell death. ORP4 silencing in non-transformed intestinal epithelial cells (IEC)-18 caused apoptosis characterized by caspase 3 and poly(ADP-ribose) polymerase processing, DNA cleavage, and JNK phosphorylation. IEC-18 transformed with oncogenic H-Ras have increased expression of ORP4L and ORP4S proteins and are resistant to the growth-inhibitory effects of ORP4 silencing. Results suggest that ORP4 promotes the survival of rapidly proliferating cells.
authors.affiliation: From the Departments of Pediatrics, Biochemistry and Molecular Biology, and.
authors.forename: Mark
authors.initials: M
authors.lastname: Charman
authors.affiliation: From the Departments of Pediatrics, Biochemistry and Molecular Biology, and.
authors.forename: Terry R
authors.initials: TR
authors.lastname: Colbourne
authors.affiliation: From the Departments of Pediatrics, Biochemistry and Molecular Biology, and.
authors.forename: Antonietta
authors.initials: A
authors.lastname: Pietrangelo
authors.affiliation: Physics and Atmospheric Sciences, Dalhousie University, Halifax, Nova Scotia B3H 4R2, Canada.
authors.forename: Laurent
authors.initials: L
authors.lastname: Kreplak
authors.affiliation: From the Departments of Pediatrics, Biochemistry and Molecular Biology, and nridgway@dal.ca.
authors.forename: Neale D
authors.initials: ND
authors.lastname: Ridgway
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D002461:Cell Line, Transformed; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D011905:Genes, ras / Q000235:genetics; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D011987:Receptors, Steroid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013261:Sterols / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism; D000098825:Oxysterol Binding Proteins
chemical_list: D018129:Phosphatidylinositol Phosphates; D010743:Phospholipids; D011987:Receptors, Steroid; D013261:Sterols; D014746:Vimentin; D000098825:Oxysterol Binding Proteins; C037178:phosphatidylinositol 4-phosphate
keywords: Apoptosis; Cholesterol; Lipid Transport; Oxysterol-binding Protein; Phosphatidylinositol 4-Phosphate; Phospholipid; RNA Interference (RNAi); Vimentin
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