Proteomic interaction profiling reveals KIFC1 as a factor involved in early targeting of F508del-CFTR to degradation

Sara Canato, João D. Santos, Ana S. Carvalho, Kerman Aloria, Margarida D. Amaral, Rune Matthiesen, André O. Falcao, Carlos M. Farinha

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Misfolded F508del-CFTR, the main molecular cause of the recessive disorder cystic fibrosis, is recognized by the endoplasmic reticulum (ER) quality control (ERQC) resulting in its retention and early degradation. The ERQC mechanisms rely mainly on molecular chaperones and on sorting motifs, whose presence and exposure determine CFTR retention or exit through the secretory pathway. Arginine-framed tripeptides (AFTs) are ER retention motifs shown to modulate CFTR retention. However, the interactions and regulatory pathways involved in this process are still largely unknown. Here, we used proteomic interaction profiling and global bioinformatic analysis to identify factors that interact differentially with F508del-CFTR and F508del-CFTR without AFTs (F508del-4RK-CFTR) as putative regulators of this specific ERQC checkpoint. Using LC–MS/MS, we identified kinesin family member C1 (KIFC1) as a stronger interactor with F508del-CFTR versus F508del-4RK-CFTR. We further validated this interaction showing that decreasing KIFC1 levels or activity stabilizes the immature form of F508del-CFTR by reducing its degradation. We conclude that the current approach is able to identify novel putative therapeutic targets that can be ultimately used to the benefit of CF patients.

Original languageEnglish
JournalCellular and Molecular Life Sciences
DOIs
Publication statusAccepted/In press - 2018

Fingerprint

Kinesin
Quality Control
Proteomics
Endoplasmic Reticulum
Arginine
Molecular Chaperones
Secretory Pathway
Computational Biology
Cystic Fibrosis
Therapeutics

Keywords

  • Arginine-framed tripeptides
  • CFTR
  • Degradation
  • Endoplasmic reticulum quality control
  • Interactome
  • Kinesins

Cite this

Canato, Sara ; Santos, João D. ; Carvalho, Ana S. ; Aloria, Kerman ; Amaral, Margarida D. ; Matthiesen, Rune ; Falcao, André O. ; Farinha, Carlos M. / Proteomic interaction profiling reveals KIFC1 as a factor involved in early targeting of F508del-CFTR to degradation. In: Cellular and Molecular Life Sciences. 2018.
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abstract = "Misfolded F508del-CFTR, the main molecular cause of the recessive disorder cystic fibrosis, is recognized by the endoplasmic reticulum (ER) quality control (ERQC) resulting in its retention and early degradation. The ERQC mechanisms rely mainly on molecular chaperones and on sorting motifs, whose presence and exposure determine CFTR retention or exit through the secretory pathway. Arginine-framed tripeptides (AFTs) are ER retention motifs shown to modulate CFTR retention. However, the interactions and regulatory pathways involved in this process are still largely unknown. Here, we used proteomic interaction profiling and global bioinformatic analysis to identify factors that interact differentially with F508del-CFTR and F508del-CFTR without AFTs (F508del-4RK-CFTR) as putative regulators of this specific ERQC checkpoint. Using LC–MS/MS, we identified kinesin family member C1 (KIFC1) as a stronger interactor with F508del-CFTR versus F508del-4RK-CFTR. We further validated this interaction showing that decreasing KIFC1 levels or activity stabilizes the immature form of F508del-CFTR by reducing its degradation. We conclude that the current approach is able to identify novel putative therapeutic targets that can be ultimately used to the benefit of CF patients.",
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Proteomic interaction profiling reveals KIFC1 as a factor involved in early targeting of F508del-CFTR to degradation. / Canato, Sara; Santos, João D.; Carvalho, Ana S.; Aloria, Kerman; Amaral, Margarida D.; Matthiesen, Rune; Falcao, André O.; Farinha, Carlos M.

In: Cellular and Molecular Life Sciences, 2018.

Research output: Contribution to journalArticle

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T1 - Proteomic interaction profiling reveals KIFC1 as a factor involved in early targeting of F508del-CFTR to degradation

AU - Canato, Sara

AU - Santos, João D.

AU - Carvalho, Ana S.

AU - Aloria, Kerman

AU - Amaral, Margarida D.

AU - Matthiesen, Rune

AU - Falcao, André O.

AU - Farinha, Carlos M.

PY - 2018

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