Congenital chloride-losing diarrhea causing mutations in the STAS domain result in misfolding and mistrafficking of SLC26A3

Michael R. Dorwart, Nikolay Shcheynikov, Jennifer M R Baker, Julie D. Forman-Kay, Shmuel Muallem, Philip J. Thomas

Research output: Contribution to journalArticle

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Abstract

Congenital chloride-losing diarrhea (CLD) is a genetic disorder causing watery stool and dehydration. Mutations in SLC26A3 (solute carrier 26 family member 3), which functions as a coupled Cl-/HCO3 - exchanger, cause CLD. SLC26A3 is a membrane protein predicted to contain 12 transmembrane-spanning α-helices and a C-terminal STAS (sulfate transporters and anti-sigma-factor) domain homologous to the bacterial anti-sigma-factor antagonists. The STAS domain is required for SLC26A3 Cl -/HCO3 - exchange function and for the activation of cystic fibrosis transmembrane conductance regulator by SLC26A3. Here we investigate the molecular mechanism(s) by which four CLD-causing mutations (ΔY526/7, I544N, I675/6ins, and G702Tins) in the STAS domain lead to disease. In a heterologous mammalian expression system biochemical, immunohistochemical, and ion transport experiments suggest that the four CLD mutations cause SLC26A3 transporter misfolding and/or mistrafficking. Expression studies with the isolated STAS domain suggest that the I675/6ins and G702Tins mutations disrupt the STAS domain directly, whereas limited proteolysis experiments suggest that the ΔY526/7 and I544N mutations affect a later step in the folding and/or trafficking pathway. The data suggest that these CLD-causing mutations cause disease by at least two distinct molecular mechanisms, both ultimately leading to loss of functional protein at the plasma membrane.

Original languageEnglish (US)
Pages (from-to)8711-8722
Number of pages12
JournalJournal of Biological Chemistry
Volume283
Issue number13
DOIs
StatePublished - Mar 28 2008

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Sigma Factor
Sulfates
Chlorides
Mutation
Diarrhea
Chloride-Bicarbonate Antiporters
Proteolysis
Cystic Fibrosis Transmembrane Conductance Regulator
Inborn Genetic Diseases
Ion Transport
Cell membranes
Dehydration
Congenital chloride diarrhea
Ion exchange
Membrane Proteins
Experiments
Chemical activation
Cell Membrane
Ions
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

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Congenital chloride-losing diarrhea causing mutations in the STAS domain result in misfolding and mistrafficking of SLC26A3. / Dorwart, Michael R.; Shcheynikov, Nikolay; Baker, Jennifer M R; Forman-Kay, Julie D.; Muallem, Shmuel; Thomas, Philip J.

In: Journal of Biological Chemistry, Vol. 283, No. 13, 28.03.2008, p. 8711-8722.

Research output: Contribution to journalArticle

Dorwart, Michael R. ; Shcheynikov, Nikolay ; Baker, Jennifer M R ; Forman-Kay, Julie D. ; Muallem, Shmuel ; Thomas, Philip J. / Congenital chloride-losing diarrhea causing mutations in the STAS domain result in misfolding and mistrafficking of SLC26A3. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 13. pp. 8711-8722.
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