PUBLICATION

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

Authors
Hartill, V.L., van de Hoek, G., Patel, M.P., Little, R., Watson, C.M., Berry, I.R., Shoemark, A., Abdelmottaleb, D., Parkes, E., Bacchelli, C., Szymanska, K., Knoers, N.V., Scambler, P.J., Ueffing, M., Boldt, K., Yates, R., Winyard, P.J., Adler, B., Moya, E., Hattingh, L., Shenoy, A., Hogg, C., Sheridan, E., Roepman, R., Norris, D., Mitchison, H.M., Giles, R.H., Johnson, C.A.
ID
ZDB-PUB-171212-14
Date
2017
Source
Human molecular genetics   27(3): 529-545 (Journal)
Registered Authors
Keywords
none
MeSH Terms
  • ATPases Associated with Diverse Cellular Activities/genetics
  • ATPases Associated with Diverse Cellular Activities/metabolism*
  • Animals
  • Carrier Proteins/genetics
  • Carrier Proteins/metabolism*
  • Cilia/metabolism*
  • Cilia/physiology
  • DNA Helicases/genetics
  • DNA Helicases/metabolism*
  • Exome Sequencing/methods
  • Female
  • Genotype
  • HEK293 Cells
  • Humans
  • Male
  • Microtubule-Associated Proteins/genetics
  • Microtubule-Associated Proteins/metabolism*
  • Mutation, Missense/genetics
  • Pedigree
  • Phenotype
  • Tumor Suppressor Proteins/genetics
  • Tumor Suppressor Proteins/metabolism
  • Zebrafish
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
PubMed
29228333 Full text @ Hum. Mol. Genet.
Abstract
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.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping