GRAM domain proteins specialize functionally distinct ER-PM contact sites in human cells

Elife. 2018 Feb 22;7:e31019. doi: 10.7554/eLife.31019.


Endoplasmic reticulum (ER) membrane contact sites (MCSs) are crucial regulatory hubs in cells, playing roles in signaling, organelle dynamics, and ion and lipid homeostasis. Previous work demonstrated that the highly conserved yeast Ltc/Lam sterol transporters localize and function at ER MCSs. Our analysis of the human family members, GRAMD1a and GRAMD2a, demonstrates that they are ER-PM MCS proteins, which mark separate regions of the plasma membrane (PM) and perform distinct functions in vivo. GRAMD2a, but not GRAMD1a, co-localizes with the E-Syt2/3 tethers at ER-PM contacts in a PIP lipid-dependent manner and pre-marks the subset of PI(4,5)P2-enriched ER-PM MCSs utilized for STIM1 recruitment. Data from an analysis of cells lacking GRAMD2a suggest that it is an organizer of ER-PM MCSs with pleiotropic functions including calcium homeostasis. Thus, our data demonstrate the existence of multiple ER-PM domains in human cells that are functionally specialized by GRAM-domain containing proteins.

Keywords: ER-PM contact sites; GRAMD proteins; PIP lipids; cell biology; cortical ER; human; membrane contact site; store operated calcium entry.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism
  • Cell Line
  • Cell Membrane / enzymology*
  • Endoplasmic Reticulum / enzymology*
  • Homeostasis
  • Humans
  • Membrane Proteins / analysis*
  • Neoplasm Proteins / analysis
  • Phosphatidylinositol Phosphates / analysis
  • Stromal Interaction Molecule 1 / analysis
  • Synaptotagmins / analysis


  • ESYT2 protein, human
  • ESYT3 protein, human
  • GRAMD1A protein, human
  • GRAMD2A protein, human
  • Membrane Proteins
  • Neoplasm Proteins
  • Phosphatidylinositol Phosphates
  • STIM1 protein, human
  • Stromal Interaction Molecule 1
  • Synaptotagmins
  • Calcium