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 Hattingh & 8 others Anil Shenoy, Claire Hogg, Eamonn Sheridan, Ronald Roepman, Dominic Norris, Hannah M. Mitchison, Rachel H. Giles, Colin A. Johnson

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Pages (from-to)529-545
Number of pages17
JournalHuman Molecular Genetics
Volume27
Issue number3
Early online date7 Dec 2017
DOIs
Publication statusPublished - 1 Feb 2018

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Dyneins
Adenosine Triphosphatases
Heart Diseases
Zebrafish
Missense Mutation
Embryonic Structures
Kartagener Syndrome
Mutation
Cilia
Virulence
Molecular Biology
Carrier Proteins
Proteins
Phenotype

Keywords

  • 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

Cite this

Hartill, V. L., van de Hoek, G., Patel, M. P., Little, R., Watson, C. M., Berry, I. R., ... Johnson, C. A. (2018). DNAAF1 links heart laterality with the AAA+ ATPase RUVBL1 and ciliary intraflagellar transport. Human Molecular Genetics, 27(3), 529-545. https://doi.org/10.1093/hmg/ddx422
Hartill, Verity L. ; van de Hoek, Glenn ; Patel, Mitali P. ; Little, Rosie ; Watson, Christopher M. ; Berry, Ian R. ; Shoemark, Amelia ; Abdelmottaleb, Dina ; Parkes, Emma ; Bacchelli, Chiara ; Szymanska, Katarzyna ; Knoers, Nine V. A. M. ; Scambler, Peter J. ; Ueffing, Marius ; Boldt, Karsten ; Yates, Robert ; Winyard, Paul J. ; Adler, Beryl ; Moya, Eduardo ; Hattingh, Louise ; Shenoy, Anil ; Hogg, Claire ; Sheridan, Eamonn ; Roepman, Ronald ; Norris, Dominic ; Mitchison, Hannah M. ; Giles, Rachel H. ; Johnson, Colin A. / DNAAF1 links heart laterality with the AAA+ ATPase RUVBL1 and ciliary intraflagellar transport. In: Human Molecular Genetics. 2018 ; Vol. 27, No. 3. pp. 529-545.
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title = "DNAAF1 links heart laterality with the AAA+ ATPase RUVBL1 and ciliary intraflagellar transport",
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.",
keywords = "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",
author = "Hartill, {Verity L.} and {van de Hoek}, Glenn and Patel, {Mitali P.} and Rosie Little and Watson, {Christopher M.} and Berry, {Ian R.} and Amelia Shoemark and Dina Abdelmottaleb and Emma Parkes and Chiara Bacchelli and Katarzyna Szymanska and Knoers, {Nine V. A. M.} and Scambler, {Peter J.} and Marius Ueffing and Karsten Boldt and Robert Yates and Winyard, {Paul J.} and Beryl Adler and Eduardo Moya and Louise Hattingh and Anil Shenoy and Claire Hogg and Eamonn Sheridan and Ronald Roepman and Dominic Norris and Mitchison, {Hannah M.} and Giles, {Rachel H.} and Johnson, {Colin A.}",
note = "This work was supported by the British Heart Foundation (Clinical Training fellowship FS/13/32/30069 to V.H.); a Sir Jules Thorn Award for Biomedical Research (JTA/09 to E.S. and C.A.J); and the Medical Research Council (grant numbers MR/K011154/1 and MR/M000532/1 to C.A.J. and MC_U142670370 to D.P.N.); an Egyptian Government Scholarship (to D.A.); and funding from the European Community's Seventh Framework Programme FP7/2009 under grant agreement no: 241955 SYSCILIA (to C.A.J., R.R. and R.H.G.) and 305608 EURenOmics (to N.V.K.) R.H.G, R.R. and N.V.K also acknowledge support from the Dutch Kidney Foundation KOUNCIL consortium CP11.18. C.M.W. acknowledges a Medical Research Council Single Cell Grant number MR/M009084/1 (funding an Illumina HiSeq3000 instrument). P.J.S. acknowledges funding from the British Heart Foundation. H.M.M. acknowledges funding from Action Medical Research (GN2101) and Great Ormond Street Hospital Children’s Charity. This research was supported by the NIHR Great Ormond Street Hospital Biomedical Research Centre (GOSH BRC). The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.",
year = "2018",
month = "2",
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language = "English",
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Hartill, VL, van de Hoek, G, Patel, MP, Little, R, Watson, CM, Berry, IR, Shoemark, A, Abdelmottaleb, D, Parkes, E, Bacchelli, C, Szymanska, K, Knoers, NVAM, Scambler, PJ, Ueffing, M, Boldt, K, Yates, R, Winyard, PJ, Adler, B, Moya, E, Hattingh, L, Shenoy, A, Hogg, C, Sheridan, E, Roepman, R, Norris, D, Mitchison, HM, Giles, RH & Johnson, CA 2018, 'DNAAF1 links heart laterality with the AAA+ ATPase RUVBL1 and ciliary intraflagellar transport', Human Molecular Genetics, vol. 27, no. 3, pp. 529-545. https://doi.org/10.1093/hmg/ddx422

DNAAF1 links heart laterality with the AAA+ ATPase RUVBL1 and ciliary intraflagellar transport. / Hartill, Verity L.; van de Hoek, Glenn; Patel, Mitali P.; Little, Rosie; Watson, Christopher M. ; Berry, Ian R.; Shoemark, Amelia; Abdelmottaleb, Dina; Parkes, Emma; Bacchelli, Chiara; Szymanska, Katarzyna; Knoers, Nine V. A. M.; Scambler, Peter J.; Ueffing, Marius; Boldt, Karsten; Yates, Robert; Winyard, Paul J.; Adler, Beryl ; Moya, Eduardo ; Hattingh, Louise; Shenoy, Anil; Hogg, Claire; Sheridan, Eamonn ; Roepman, Ronald; Norris, Dominic; Mitchison, Hannah M. (Lead / Corresponding author); Giles, Rachel H. (Lead / Corresponding author); Johnson, Colin A. (Lead / Corresponding author).

In: Human Molecular Genetics, Vol. 27, No. 3, 01.02.2018, p. 529-545.

Research output: Contribution to journalArticle

TY - JOUR

T1 - DNAAF1 links heart laterality with the AAA+ ATPase RUVBL1 and ciliary intraflagellar transport

AU - Hartill, Verity L.

AU - van de Hoek, Glenn

AU - Patel, Mitali P.

AU - Little, Rosie

AU - Watson, Christopher M.

AU - Berry, Ian R.

AU - Shoemark, Amelia

AU - Abdelmottaleb, Dina

AU - Parkes, Emma

AU - Bacchelli, Chiara

AU - Szymanska, Katarzyna

AU - Knoers, Nine V. A. M.

AU - Scambler, Peter J.

AU - Ueffing, Marius

AU - Boldt, Karsten

AU - Yates, Robert

AU - Winyard, Paul J.

AU - Adler, Beryl

AU - Moya, Eduardo

AU - Hattingh, Louise

AU - Shenoy, Anil

AU - Hogg, Claire

AU - Sheridan, Eamonn

AU - Roepman, Ronald

AU - Norris, Dominic

AU - Mitchison, Hannah M.

AU - Giles, Rachel H.

AU - Johnson, Colin A.

N1 - This work was supported by the British Heart Foundation (Clinical Training fellowship FS/13/32/30069 to V.H.); a Sir Jules Thorn Award for Biomedical Research (JTA/09 to E.S. and C.A.J); and the Medical Research Council (grant numbers MR/K011154/1 and MR/M000532/1 to C.A.J. and MC_U142670370 to D.P.N.); an Egyptian Government Scholarship (to D.A.); and funding from the European Community's Seventh Framework Programme FP7/2009 under grant agreement no: 241955 SYSCILIA (to C.A.J., R.R. and R.H.G.) and 305608 EURenOmics (to N.V.K.) R.H.G, R.R. and N.V.K also acknowledge support from the Dutch Kidney Foundation KOUNCIL consortium CP11.18. C.M.W. acknowledges a Medical Research Council Single Cell Grant number MR/M009084/1 (funding an Illumina HiSeq3000 instrument). P.J.S. acknowledges funding from the British Heart Foundation. H.M.M. acknowledges funding from Action Medical Research (GN2101) and Great Ormond Street Hospital Children’s Charity. This research was supported by the NIHR Great Ormond Street Hospital Biomedical Research Centre (GOSH BRC). The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

PY - 2018/2/1

Y1 - 2018/2/1

N2 - 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.

AB - 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.

KW - ATPases Associated with Diverse Cellular Activities/genetics

KW - Animals

KW - Carrier Proteins/genetics

KW - Cilia/metabolism

KW - DNA Helicases/genetics

KW - Female

KW - Genotype

KW - HEK293 Cells

KW - Humans

KW - Male

KW - Microtubule-Associated Proteins/genetics

KW - Mutation, Missense/genetics

KW - Pedigree

KW - Phenotype

KW - Tumor Suppressor Proteins/genetics

KW - Whole Exome Sequencing/methods

KW - Zebrafish

KW - Zebrafish Proteins/genetics

U2 - 10.1093/hmg/ddx422

DO - 10.1093/hmg/ddx422

M3 - Article

VL - 27

SP - 529

EP - 545

JO - Human Molecular Genetics

JF - Human Molecular Genetics

SN - 0964-6906

IS - 3

ER -

Hartill VL, van de Hoek G, Patel MP, Little R, Watson CM, Berry IR et al. DNAAF1 links heart laterality with the AAA+ ATPase RUVBL1 and ciliary intraflagellar transport. Human Molecular Genetics. 2018 Feb 1;27(3):529-545. https://doi.org/10.1093/hmg/ddx422