Abstract
Leber congenital amaurosis (LCA) causes blindness or severe visual impairment at or within a few months of birth. Here we show, using homozygosity mapping, that the LCA5 gene on chromosome 6q14, which encodes the previously unknown ciliary protein lebercilin, is associated with this disease. We detected homozygous nonsense and frameshift mutations in LCA5 in five families affected with LCA. In a sixth family, the LCA5 transcript was completely absent. LCA5 is expressed widely throughout development, although the phenotype in affected individuals is limited to the eye. Lebercilin localizes to the connecting cilia of photoreceptors and to the microtubules, centrioles and primary cilia of cultured mammalian cells. Using tandem affinity purification, we identified 24 proteins that link lebercilin to centrosomal and ciliary functions. Members of this interactome represent candidate genes for LCA and other ciliopathies. Our findings emphasize the emerging role of disrupted ciliary processes in the molecular pathogenesis of LCA.
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Acknowledgements
We thank the LCA families for their participation; H. Brunner, C. Johnson, N. Knoers and H. Kremer for discussions; C.J. Gloeckner for the TAPe constructs; T. Goldmann, E. Sehn, J. Hehir-Kwa, I. Janssen, K. Voesenek, A. Schumacher and S. Schöffmann for technical assistance; K. Klima, R. Pigeon and C. Robert for organizing the clinical data from affected individuals; S. Yzer, L.I. van den Born, S. Kohl, B. Wissinger, E. de Baere, B.P. Leroy, W. Bergen, K. Rohrschneider and C.B. Hoyng for sharing patient samples; and the Marshfield Mammalian Genotyping Service for carrying out genotyping in the Pakistani families. This research was supported by grants from The Netherlands Organisation for Scientific Research (916.56.160 to A.I.d.H.); the Foundation Fighting Blindness USA (BR-GE-0606-0349-RAD to A.I.d.H.); the Dutch Kidney Foundation (C04.2112 to R.R.); Landelijke Stichting voor Blinden en Slechtzienden (to A.I.d.H. and F.P.M.C.); Algemene Nederlandse Vereniging ter Voorkoming van Blindheid (to A.I.d.H., R.R. and F.P.M.C); Rotterdamse Vereniging Blindenbelangen (to R.R. and F.P.M.C.); Stichting Blindenhulp (to R.R. and F.P.M.C.); Stichting OOG (to R.R. and F.P.M.C.); the British Retinitis Pigmentosa Society (GR552 to R.R.); the European Union 6th Framework RETNET (MRTNCT-2003-504003 to F.P.M.C. and M.U.), EVI-GENORET (LSHG-CT-2005 512036 to M.U., F.P.M.C. and R.R.) and INTERACTION PROTEOME (LSHG-CT-2003-505520 to M.U.); the Wellcome Trust (061682 and 073477 to C.F.I. and M.D.M., and 068579 to M.E.C.); Yorkshire Eye Research (006 to C.F.I.); the Foundation Fighting Blindness Canada (to R.K.K. and F.P.M.C.); the Fonds de la Recherche en Santé Québec; TD Financial Group (to R.K.K.); Foundation of Retinal Research; The Grousbeck Foundation; The Edel and Krieble Funds; the Ort Family Foundation (to I.H.M., S.D. and R.K.K.); Pro Retina Germany (to M.U. and R.R.); Deutsche Forschungsgemeinschaft (Wo 548-6 to U.W.), FAUN-Stiftung (to U.W.) and Forschung contra Blindheit (to U.W.).
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Supplementary information
Supplementary Fig. 1
Sequences of LCA5 mutations identified in LCA families. (PDF 26 kb)
Supplementary Fig. 2
Evolutionary conservation of lebercilin and C21ORF13 proteins. (PDF 21 kb)
Supplementary Fig. 3
Expression of the LCA5 gene in human tissues and mammalian cell lines, and detection of lebercilin in mouse tissues. (PDF 467 kb)
Supplementary Fig. 4
Expression of the recombinant lebercilin proteins in ARPE-19 and COS-1 cells. (PDF 328 kb)
Supplementary Fig. 5
Expression of the TAPe constructs and immunoprecipitation of lebercilin. (PDF 422 kb)
Supplementary Table 1
Refinement of the LCA5 interval in three Pakistani LCA families. (PDF 49 kb)
Supplementary Table 2
Homozygous chromosomal regions in patients 27240 and 28609. (PDF 46 kb)
Supplementary Table 3
Protein/peptide summaries of LC-MSMS analysis of tandem affinity-purified lebercilin protein complexes. (PDF 489 kb)
Supplementary Table 4
Primer sequences for amplification of the exons and splice junctions of the LCA5 gene. (PDF 48 kb)
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den Hollander, A., Koenekoop, R., Mohamed, M. et al. Mutations in LCA5, encoding the ciliary protein lebercilin, cause Leber congenital amaurosis. Nat Genet 39, 889–895 (2007). https://doi.org/10.1038/ng2066
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DOI: https://doi.org/10.1038/ng2066
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