Binding of the erlin1/2 complex to the third intralumenal loop of IP3R1 triggers its ubiquitin-proteasomal degradation

J Biol Chem. 2022 Jun;298(6):102026. doi: 10.1016/j.jbc.2022.102026. Epub 2022 May 11.

Abstract

Long-term activation of inositol 1,4,5-trisphosphate receptors (IP3Rs) leads to their degradation by the ubiquitin-proteasome pathway. The first and rate-limiting step in this process is thought to be the association of conformationally active IP3Rs with the erlin1/2 complex, an endoplasmic reticulum-located oligomer of erlin1 and erlin2 that recruits the E3 ubiquitin ligase RNF170, but the molecular determinants of this interaction remain unknown. Here, through mutation of IP3R1, we show that the erlin1/2 complex interacts with the IP3R1 intralumenal loop 3 (IL3), the loop between transmembrane (TM) helices 5 and 6, and in particular, with a region close to TM5, since mutation of amino acids D-2471 and R-2472 can specifically block erlin1/2 complex association. Surprisingly, we found that additional mutations in IL3 immediately adjacent to TM5 (e.g., D2465N) almost completely abolish IP3R1 Ca2+ channel activity, indicating that the integrity of this region is critical to IP3R1 function. Finally, we demonstrate that inhibition of the ubiquitin-activating enzyme UBE1 by the small-molecule inhibitor TAK-243 completely blocked IP3R1 ubiquitination and degradation without altering erlin1/2 complex association, confirming that association of the erlin1/2 complex is the primary event that initiates IP3R1 processing and that IP3R1 ubiquitination mediates IP3R1 degradation. Overall, these data localize the erlin1/2 complex-binding site on IP3R1 to IL3 and show that the region immediately adjacent to TM5 is key to the events that facilitate channel opening.

Keywords: calcium channel; endoplasmic reticulum–associated degradation; erlin1/2 complex; inositol trisphosphate receptor; protein–protein interaction; ubiquitination.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Endoplasmic Reticulum* / genetics
  • Endoplasmic Reticulum* / metabolism
  • Inositol 1,4,5-Trisphosphate Receptors* / genetics
  • Inositol 1,4,5-Trisphosphate Receptors* / metabolism
  • Membrane Proteins* / genetics
  • Membrane Proteins* / metabolism
  • Mice
  • Ubiquitin* / metabolism
  • Ubiquitination

Substances

  • Inositol 1,4,5-Trisphosphate Receptors
  • Itpr1 protein, mouse
  • Membrane Proteins
  • Ubiquitin
  • erlin-1 protein, mouse
  • erlin-2 protein, mouse