DNAAF1 links heart laterality with the AAA+ ATPase RUVBL1 and ciliary intraflagellar transport

Verity L. Hartill, Glenn van de Hoek, Mitali P. Patel, Rosie Little, Christopher M. Watson, Ian R. Berry, Amelia Shoemark, Dina Abdelmottaleb, Emma Parkes, Chiara Bacchelli, Katarzyna Szymanska, Nine V. A. M. Knoers, Peter J. Scambler, Marius Ueffing, Karsten Boldt, Robert Yates, Paul J. Winyard, Beryl Adler, Eduardo Moya, Louise HattinghAnil Shenoy, Claire Hogg, Eamonn Sheridan, Ronald Roepman, Dominic Norris, Hannah M. Mitchison (Lead / Corresponding author), Rachel H. Giles (Lead / Corresponding author), Colin A. Johnson (Lead / Corresponding author)

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25 Citations (Scopus)
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DNAAF1 (LRRC50) is a cytoplasmic protein required for dynein heavy chain assembly and cilia motility, and DNAAF1 mutations cause primary ciliary dyskinesia (PCD; MIM 613193). We describe four families with DNAAF1 mutations and complex congenital heart disease (CHD). In three families, all affected individuals have typical PCD phenotypes. However, an additional family demonstrates isolated CHD (heterotaxy) in two affected siblings, but no clinical evidence of PCD. We identified a homozygous DNAAF1 missense mutation, p.Leu191Phe, as causative for heterotaxy in this family. Genetic complementation in dnaaf1-null zebrafish embryos demonstrated the rescue of normal heart looping with wild-type human DNAAF1, but not the p.Leu191Phe variant, supporting the conserved pathogenicity of this DNAAF1 missense mutation. This observation points to a phenotypic continuum between CHD and PCD, providing new insights into the pathogenesis of isolated CHD. In further investigations of the function of DNAAF1 in dynein arm assembly, we identified interactions with members of a putative dynein arm assembly complex. These include the ciliary intraflagellar transport protein IFT88 and the AAA + (ATPases Associated with various cellular Activities) family proteins RUVBL1 (Pontin) and RUVBL2 (Reptin). Co-localization studies support these findings, with the loss of RUVBL1 perturbing the co-localization of DNAAF1 with IFT88. We show that RUVBL1 orthologues have an asymmetric left-sided distribution at both the mouse embryonic node and the Kupffer's vesicle in zebrafish embryos, with the latter asymmetry dependent on DNAAF1. These results suggest that DNAAF1-RUVBL1 biochemical and genetic interactions have a novel functional role in symmetry breaking and cardiac development.

Original languageEnglish
Pages (from-to)529-545
Number of pages17
JournalHuman Molecular Genetics
Issue number3
Early online date7 Dec 2017
Publication statusPublished - 1 Feb 2018


  • ATPases Associated with Diverse Cellular Activities/genetics
  • Animals
  • Carrier Proteins/genetics
  • Cilia/metabolism
  • DNA Helicases/genetics
  • Female
  • Genotype
  • HEK293 Cells
  • Humans
  • Male
  • Microtubule-Associated Proteins/genetics
  • Mutation, Missense/genetics
  • Pedigree
  • Phenotype
  • Tumor Suppressor Proteins/genetics
  • Whole Exome Sequencing/methods
  • Zebrafish
  • Zebrafish Proteins/genetics

ASJC Scopus subject areas

  • Genetics(clinical)
  • Genetics
  • Molecular Biology


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