Chaperone-Independent Peripheral Quality Control of CFTR by RFFL E3 Ligase

Tsukasa Okiyoneda, Guido Veit, Ryohei Sakai, Misaki Aki, Takeshi Fujihara, Momoko Higashi, Seiko Susuki-Miyata, Masanori Miyata, Norihito Fukuda, Akihiko Yoshida, Haijin Xu, Pirjo M. Apaja, Gergely L. Lukacs

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

The peripheral protein quality control (QC) system removes non-native membrane proteins, including ΔF508-CFTR, the most common CFTR mutant in cystic fibrosis (CF), from the plasma membrane (PM) for lysosomal degradation by ubiquitination. It remains unclear how unfolded membrane proteins are recognized and targeted for ubiquitination and how they are removed from the apical PM. Using comprehensive siRNA screens, we identified RFFL, an E3 ubiquitin (Ub) ligase that directly and selectively recognizes unfolded ΔF508-CFTR through its disordered regions. RFFL retrieves the unfolded CFTR from the PM for lysosomal degradation by chaperone-independent K63-linked poly-ubiquitination. RFFL ablation enhanced the functional expression of cell-surface ΔF508-CFTR in the presence of folding corrector molecules, and this effect was further improved by inhibiting the Hsc70-dependent ubiquitination machinery. We propose that multiple peripheral QC mechanisms evolved to dispose of non-native PM proteins and to preserve cellular proteostasis, even at the cost of eliminating partially functional polypeptides. Okiyoneda et al. investigate how ubiquitination targets ΔF508-CFTR, the most common CFTR mutant found in cystic fibrosis, at the cell surface. The authors show that RFFL, a ubiquitin ligase that directly and selectively recognizes unfolded ΔF508-CFTR through its disordered regions, promotes K63-linked poly-ubiquitination of CFTR to stimulate lysosomal degradation.

Original languageEnglish
Pages (from-to)694-708.e7
JournalDevelopmental Cell
Volume44
Issue number6
DOIs
Publication statusPublished - 26 Mar 2018
Externally publishedYes

Keywords

  • CFTR
  • cystic fibrosis
  • endosome
  • protein quality control
  • ubiquitination

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Developmental Biology
  • Cell Biology

Cite this