Alcohol drives S-nitrosylation and redox activation of protein phosphatase 1, causing bovine airway cilia dysfunction

Am J Physiol Lung Cell Mol Physiol. 2017 Mar 1;312(3):L432-L439. doi: 10.1152/ajplung.00513.2016. Epub 2017 Jan 6.

Abstract

Individuals with alcohol (ethanol)-use disorders are at increased risk for lung infections, in part, due to defective mucociliary clearance driven by motile cilia in the airways. We recently reported that isolated, demembranated bovine cilia (axonemes) are capable of producing nitric oxide (NO) when exposed to biologically relevant concentrations of alcohol. This increased presence of NO can lead to protein S-nitrosylation, a posttranslational modification signaling mechanism involving reversible adduction of nitrosonium cations or NO to thiolate or thiyl radicals, respectively, of proteins forming S-nitrosothiols (SNOs). We quantified and compared SNO content between isolated, demembranated axonemes extracted from bovine tracheae, with or without in situ alcohol exposure (100 mM × 24 h). We demonstrate that relevant concentrations of alcohol exposure shift the S-nitrosylation status of key cilia regulatory proteins, including 20-fold increases in S-nitrosylation of proteins that include protein phosphatase 1 (PP1). With the use of an ATP-reactivated axoneme motility system, we demonstrate that alcohol-driven S-nitrosylation of PP1 is associated with PP1 activation and dysfunction of axoneme motility. These new data demonstrate that alcohol can shift the S-nitrothiol balance at the level of the cilia organelle and highlight S-nitrosylation as a novel signaling mechanism to regulate PP1 and cilia motility.

Keywords: S-nitrosylation; alcohol; cilia; redox regulation.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axoneme / drug effects
  • Axoneme / metabolism
  • Cattle
  • Cilia / drug effects
  • Cilia / pathology*
  • Ethanol / toxicity*
  • Nitrosation
  • Oxidation-Reduction / drug effects
  • Protein Phosphatase 1 / metabolism*
  • Proteome / metabolism
  • Trachea / drug effects
  • Trachea / pathology*
  • Trachea / physiopathology*

Substances

  • Proteome
  • Ethanol
  • Protein Phosphatase 1