SYVN1, NEDD8, and FBXO2 Proteins Regulate ΔF508 Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Ubiquitin-mediated Proteasomal Degradation

Shyam Ramachandran; Samantha R Osterhaus; Kalpaj R Parekh; Ashley M Jacobi; Mark A Behlke; Paul B McCray Jr

doi:10.1074/jbc.M116.754283

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SYVN1, NEDD8, and FBXO2 Proteins Regulate ΔF508 Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Ubiquitin-mediated Proteasomal Degradation

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SYVN1, NEDD8, and FBXO2 Proteins Regulate ΔF508 Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Ubiquitin-mediated Proteasomal Degradation

Shyam Ramachandran, Samantha R Osterhaus, Kalpaj R Parekh, Ashley M Jacobi, Mark A Behlke and Paul B McCray Jr

The Journal of biological chemistry, Vol.291(49), pp.25489-25504

12/02/2016

DOI: 10.1074/jbc.M116.754283

PMCID: PMC5207249

PMID: 27756846

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Abstract

We previously reported that delivery of a microRNA-138 mimic or siRNA against SIN3A to cultured cystic fibrosis (ΔF508/ΔF508) airway epithelia partially restored ΔF508-cystic fibrosis transmembrane conductance regulator (CFTR)-mediated cAMP-stimulated Cl conductance. We hypothesized that dissecting this microRNA-138/SIN3A-regulated gene network would identify individual proteins contributing to the rescue of ΔF508-CFTR function. Among the genes in the network, we rigorously validated candidates using functional CFTR maturation and electrolyte transport assays in polarized airway epithelia. We found that depletion of the ubiquitin ligase SYVN1, the ubiquitin/proteasome system regulator NEDD8, or the F-box protein FBXO2 partially restored ΔF508-CFTR-mediated Cl transport in primary cultures of human cystic fibrosis airway epithelia. Moreover, knockdown of SYVN1, NEDD8, or FBXO2 in combination with corrector compound 18 further potentiated rescue of ΔF508-CFTR-mediated Cl conductance. This study provides new knowledge of the CFTR biosynthetic pathway. It suggests that SYVN1 and FBXO2 represent two distinct multiprotein complexes that may degrade ΔF508-CFTR in airway epithelia and identifies a new role for NEDD8 in regulating ΔF508-CFTR ubiquitination.

Sequence Deletion

Epithelial Cells - metabolism

Humans

Ubiquitin - metabolism

Gene Knockdown Techniques

Ubiquitination

Proteolysis

Cell Cycle Proteins - genetics

Ubiquitins - metabolism

NEDD8 Protein

Amino Acid Sequence

F-Box Proteins - metabolism

Cell Cycle Proteins - metabolism

Ubiquitin-Protein Ligases - metabolism

Epithelial Cells - pathology

Cystic Fibrosis Transmembrane Conductance Regulator - metabolism

Ubiquitin - genetics

Nerve Tissue Proteins - genetics

Nerve Tissue Proteins - metabolism

Proteasome Endopeptidase Complex - genetics

Cystic Fibrosis Transmembrane Conductance Regulator - genetics

Respiratory Mucosa - physiology

HeLa Cells

Proteasome Endopeptidase Complex - metabolism

Respiratory Mucosa - metabolism

Ubiquitin-Protein Ligases - genetics

F-Box Proteins - genetics

Ion Transport - genetics

Details

Title: Subtitle: SYVN1, NEDD8, and FBXO2 Proteins Regulate ΔF508 Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Ubiquitin-mediated Proteasomal Degradation
Creators: Shyam Ramachandran - From the Department of Pediatrics, Pappajohn Biomedical Institute, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, Iowa 52242 and
Samantha R Osterhaus - From the Department of Pediatrics, Pappajohn Biomedical Institute, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, Iowa 52242 and
Kalpaj R Parekh - From the Department of Pediatrics, Pappajohn Biomedical Institute, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, Iowa 52242 and
Ashley M Jacobi - Integrated DNA Technologies, Coralville, Iowa 52241
Mark A Behlke - Integrated DNA Technologies, Coralville, Iowa 52241
Paul B McCray Jr - From the Department of Pediatrics, Pappajohn Biomedical Institute, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, Iowa 52242 and paul-mccray@uiowa.edu
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.291(49), pp.25489-25504
DOI: 10.1074/jbc.M116.754283
PMID: 27756846
PMCID: PMC5207249
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: P01 HL091842 / NHLBI NIH HHS R01 HL118000 / NHLBI NIH HHS P01 HL051670 / NHLBI NIH HHS R21 HL104337 / NHLBI NIH HHS P30 DK054759 / NIDDK NIH HHS
Language: English
Date published: 12/02/2016
Academic Unit: Microbiology and Immunology; Pulmonary Medicine; Anatomy and Cell Biology; Stead Family Department of Pediatrics; Surgery; Cardiothoracic Surgery; Internal Medicine
Record Identifier: 9984025313402771

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