title: Alcohol-induced ciliary dysfunction targets the outer dynein arm.
abstract: Alcohol abuse results in an increased incidence of pulmonary infection, in part attributable to impaired mucociliary clearance. Analysis of motility in mammalian airway cilia has revealed that alcohol impacts the ciliary dynein motors by a mechanism involving altered axonemal protein phosphorylation. Given the highly conserved nature of cilia, it is likely that the mechanisms for alcohol-induced ciliary dysfunction (AICD) are conserved. Thus we utilized the experimental advantages offered by the model organism, Chlamydomonas, to determine the precise effects of alcohol on ciliary dynein activity and identify axonemal phosphoproteins that are altered by alcohol exposure. Analysis of live cells or reactivated cell models showed that alcohol significantly inhibits ciliary motility in Chlamydomonas via a mechanism that is part of the axonemal structure. Taking advantage of informative mutant cells, we found that alcohol impacts the activity of the outer dynein arm. Consistent with this finding, alcohol exposure results in a significant reduction in ciliary beat frequency, a parameter of ciliary movement that requires normal outer dynein arm function. Using mutants that lack specific heavy-chain motor domains, we have determined that alcohol impacts the β- and γ-heavy chains of the outer dynein arm. Furthermore, using a phospho-threonine-specific antibody, we determined that the phosphorylation state of DCC1 of the outer dynein arm-docking complex is altered in the presence of alcohol, and its phosphorylation correlates with AICD. These results demonstrate that alcohol targets specific outer dynein arm components and suggest that DCC1 is part of an alcohol-sensitive mechanism that controls outer dynein arm activity.
authors.affiliation: University of Mississippi Medical Center, Department of Biochemistry, Jackson, Mississippi;
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: University of Nebraska Medical Center, Department of Internal Medicine, Division of Pulmonary, Critical Care, Sleep and Allergy, Omaha, Nebraska; and.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Pavlik
authors.affiliation: Emory University School of Medicine, Department of Cell Biology, Atlanta, Georgia.
authors.forename: Laura
authors.initials: L
authors.lastname: Fox
authors.affiliation: University of Mississippi Medical Center, Department of Biochemistry, Jackson, Mississippi;
authors.forename: Chasity
authors.initials: C
authors.lastname: Scarbrough
authors.affiliation: Emory University School of Medicine, Department of Cell Biology, Atlanta, Georgia.
authors.forename: Winfield S
authors.initials: WS
authors.lastname: Sale
authors.affiliation: University of Nebraska Medical Center, Department of Internal Medicine, Division of Pulmonary, Critical Care, Sleep and Allergy, Omaha, Nebraska; and.
authors.forename: Joseph H
authors.initials: JH
authors.lastname: Sisson
authors.affiliation: University of Mississippi Medical Center, Department of Biochemistry, Jackson, Mississippi; mwirschell@umc.edu.
authors.forename: Maureen
authors.initials: M
authors.lastname: Wirschell
mesh_terms: D054468:Axoneme / Q000235:genetics / Q000378:metabolism*; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D002696:Chlamydomonas / Q000235:genetics / Q000378:metabolism*; D002923:Cilia / Q000235:genetics / Q000378:metabolism; D004398:Dyneins / Q000235:genetics / Q000378:metabolism*; D000431:Ethanol / Q000494:pharmacology*; D009154:Mutation
chemical_list: D002492:Central Nervous System Depressants; D000431:Ethanol; D004398:Dyneins
keywords: alcohol; cilia; dynein; ethanol