Orexin/hypocretin activates mTOR complex 1 (mTORC1) via an Erk/Akt-independent and calcium-stimulated lysosome v-ATPase pathway

J Biol Chem. 2014 Nov 14;289(46):31950-31959. doi: 10.1074/jbc.M114.600015. Epub 2014 Oct 2.

Abstract

The lack of the neuropeptide orexin, also known as hypocretin, results in narcolepsy, a chronic sleep disorder characterized by frequent sleep/cataplexy attacks and rapid eye movement sleep abnormalities. However, the downstream pathways of orexin signaling are not clearly understood. Here, we show that orexin activates the mTOR pathway, a central regulator of cell growth and metabolism, in the mouse brain and multiple recombinant cell lines that express the G protein-coupled receptors (GPCRs), orexin 1 receptor (OX1R) or orexin 2 receptor (OX2R). This orexin/GPCR-stimulated mTOR activation is sensitive to rapamycin, an inhibitor of mTOR complex 1 (mTORC1) but is independent of two well known mTORC1 activators, Erk and Akt. Rather, our studies indicate that orexin activates mTORC1 via extracellular calcium influx and the lysosome pathway involving v-ATPase and Rag GTPases. Moreover, a cytoplasmic calcium transient is sufficient to mimic orexin/GPCR signaling to mTORC1 activation in a v-ATPase-dependent manner. Together, our studies suggest that the mTORC1 pathway functions downstream of orexin/GPCR signaling, which plays a crucial role in many physiological and metabolic processes.

Keywords: Calcium; G Protein-coupled Receptor (GPCR); Lysosome; Neuropeptide; OX1R Or OX2R; Orexin; Rag GTPase; Vacuolar ATPase; mTOR Complex (mTORC).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / metabolism*
  • Calcium / metabolism
  • Cell Line
  • Cytoplasm / metabolism
  • Enzyme Activation
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Gene Expression Regulation, Enzymologic
  • HEK293 Cells
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Lysosomes / metabolism
  • Mechanistic Target of Rapamycin Complex 1
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Multiprotein Complexes / metabolism*
  • Neuropeptides / metabolism*
  • Orexin Receptors / metabolism*
  • Orexins
  • Proto-Oncogene Proteins c-akt / metabolism
  • RNA Interference
  • Signal Transduction
  • TOR Serine-Threonine Kinases / metabolism*
  • Vacuolar Proton-Translocating ATPases / metabolism*

Substances

  • Cd200r1 protein, mouse
  • HCRTR1 protein, human
  • HCRTR2 protein, human
  • Hcrtr1 protein, mouse
  • Intracellular Signaling Peptides and Proteins
  • Multiprotein Complexes
  • Neuropeptides
  • Orexin Receptors
  • Orexins
  • AKT1 protein, human
  • Akt1 protein, mouse
  • Mechanistic Target of Rapamycin Complex 1
  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases
  • Extracellular Signal-Regulated MAP Kinases
  • Vacuolar Proton-Translocating ATPases
  • Calcium