Oxysterol-binding protein-related protein 5 (ORP5) promotes cell proliferation by activation of mTORC1 signaling

J Biol Chem. 2018 Mar 9;293(10):3806-3818. doi: 10.1074/jbc.RA117.001558. Epub 2018 Jan 22.

Abstract

Oxysterol-binding protein (OSBP) and OSBP-related proteins (ORPs) constitute a large family of proteins that mainly function in lipid transport and sensing. ORP5 is an endoplasmic reticulum (ER)-anchored protein implicated in lipid transfer at the contact sites between the ER and other membranes. Recent studies indicate that ORP5 is also involved in cancer cell invasion and tumor progression. However, the molecular mechanism underlying ORP5's involvement in cancer is unclear. Here, we report that ORP5 promotes cell proliferation and motility of HeLa cells, an effect that depends on its functional OSBP-related domain (ORD). We also found that ORP5 depletion or substitutions of key residues located within ORP5-ORD and responsible for interactions with lipids interfered with cell proliferation, migration, and invasion. ORP5 interacted with the protein mechanistic target of rapamycin (mTOR), and this interaction also required ORP5-ORD. Of note, whereas ORP5 overexpression induced mTOR complex 1 (mTORC1) activity, ORP5 down-regulation had the opposite effect. Finally, ORP5-depleted cells exhibited impaired mTOR localization to lysosomes, which may have accounted for the blunted mTORC1 activation. Together, our results suggest that ORP5 expression is positively correlated with mTORC1 signaling and that ORP5 stimulates cell proliferation, at least in part, by activating mTORC1.

Keywords: ORP5; ORP8; OSBP; cancer; cell proliferation; cell signaling; lipids; mTOR complex (mTORC); mammalian target of rapamycin (mTOR); phosphoinositide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Cell Line, Tumor
  • Cell Movement
  • Cell Proliferation
  • Enzyme Activation
  • Gene Deletion
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Lysosomes / enzymology
  • Lysosomes / metabolism*
  • Lysosomes / pathology
  • Mechanistic Target of Rapamycin Complex 1 / agonists*
  • Mechanistic Target of Rapamycin Complex 1 / metabolism
  • Neoplasm Invasiveness
  • Neoplasm Proteins / antagonists & inhibitors
  • Neoplasm Proteins / chemistry
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Neoplasms / enzymology
  • Neoplasms / metabolism*
  • Neoplasms / pathology
  • Point Mutation
  • Protein Interaction Domains and Motifs
  • Protein Transport
  • RNA Interference
  • Receptors, Steroid / antagonists & inhibitors
  • Receptors, Steroid / chemistry
  • Receptors, Steroid / genetics
  • Receptors, Steroid / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction*
  • TOR Serine-Threonine Kinases / metabolism*
  • Up-Regulation*

Substances

  • Neoplasm Proteins
  • Receptors, Steroid
  • Recombinant Fusion Proteins
  • oxysterol binding protein
  • Green Fluorescent Proteins
  • MTOR protein, human
  • Mechanistic Target of Rapamycin Complex 1
  • TOR Serine-Threonine Kinases