Regulatory R region of the CFTR chloride channel is a dynamic integrator of phospho-dependent intra- and intermolecular interactions

Zoltan Bozoky, Mickael Krzeminski, Ranjith Muhandiram, James R. Birtley, Ateeq Al-Zahrani, Philip J. Thomas, Raymond A. Frizzell, Robert C. Ford, Julie D. Forman-Kay

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Abstract

Intrinsically disordered proteins play crucial roles in regulatory processes and often function as protein interaction hubs. Here, we present a detailed characterization of a full-length disordered hub protein region involved in multiple dynamic complexes. We performed NMR, CD, and fluorescence binding studies on the nonphosphorylated and highly PKA-phosphorylated human cystic fibrosis transmembrane conductance regulator (CFTR) regulatory region, a ∼200-residue disordered segment involved in phosphorylation- dependent regulation of channel trafficking and gating. Our data provide evidence for dynamic, phosphorylation-dependent, multisite interactions of various segments of the regulatory region for its intra- and intermolecular partners, including the CFTR nucleotide binding domains 1 and 2, a 42-residue peptide from the C terminus of CFTR, the SLC26A3 sulphate transporter and antisigma factor antagonist (STAS) domain, and 14-3-3β. Because of its large number of binding partners, multivalent binding of individually weak sites facilitates rapid exchange between free and bound states to allow the regulatory region to engage with different partners and generate a graded or rheostat-like response to phosphorylation. Our results enrich the understanding of how disordered binding segments interact with multiple targets. We present structural models consistent with our data that illustrate this dynamic aspect of phospho-regulation of CFTR by the disordered regulatory region.

Original languageEnglish (US)
JournalProceedings of the National Academy of Sciences of the United States of America
Volume110
Issue number47
DOIs
StatePublished - Nov 19 2013

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Cystic Fibrosis Transmembrane Conductance Regulator
Chloride Channels
Nucleic Acid Regulatory Sequences
Phosphorylation
Intrinsically Disordered Proteins
Structural Models
Sulfates
Proteins
Nucleotides
Fluorescence

ASJC Scopus subject areas

  • General

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Regulatory R region of the CFTR chloride channel is a dynamic integrator of phospho-dependent intra- and intermolecular interactions. / Bozoky, Zoltan; Krzeminski, Mickael; Muhandiram, Ranjith; Birtley, James R.; Al-Zahrani, Ateeq; Thomas, Philip J.; Frizzell, Raymond A.; Ford, Robert C.; Forman-Kay, Julie D.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 110, No. 47, 19.11.2013.

Research output: Contribution to journalArticle

Bozoky, Zoltan ; Krzeminski, Mickael ; Muhandiram, Ranjith ; Birtley, James R. ; Al-Zahrani, Ateeq ; Thomas, Philip J. ; Frizzell, Raymond A. ; Ford, Robert C. ; Forman-Kay, Julie D. / Regulatory R region of the CFTR chloride channel is a dynamic integrator of phospho-dependent intra- and intermolecular interactions. In: Proceedings of the National Academy of Sciences of the United States of America. 2013 ; Vol. 110, No. 47.
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