The kinetochore protein, CENPF, is mutated in human ciliopathy and microcephaly phenotypes

• Published in: Journal of medical genetics Vol. 52; no. 3; pp. 147 - 156
• Main Authors: Waters, Aoife M, Asfahani, Rowan, Carroll, Paula, Bicknell, Louise, Lescai, Francesco, Bright, Alison, Chanudet, Estelle, Brooks, Anthony, Christou-Savina, Sonja, Osman, Guled, Walsh, Patrick, Bacchelli, Chiara, Chapgier, Ariane, Vernay, Bertrand, Bader, David M, Deshpande, Charu, O’ Sullivan, Mary, Ocaka, Louise, Stanescu, Horia, Stewart, Helen S, Hildebrandt, Friedhelm, Otto, Edgar, Johnson, Colin A, Szymanska, Katarzyna, Katsanis, Nicholas, Davis, Erica, Kleta, Robert, Hubank, Mike, Doxsey, Stephen, Jackson, Andrew, Stupka, Elia, Winey, Mark, Beales, Philip L
• Format: Journal Article
• Language: English
• Published: England BMJ Publishing Group LTD 01.03.2015; BMJ Publishing Group
• Series: Original article
• Subjects: Animals; Antibodies; Centrioles - genetics; Chromosomal Proteins, Non-Histone - genetics; Cilia - genetics; Cilia - pathology; Developmental Defects; Exome - genetics; Female; Fetus; Fetuses; Fibroblasts; Genes; Genetics, Medical; Genomes; Genomics; HEK293 Cells; High-Throughput Nucleotide Sequencing; Humans; Male; Mice; Microcephaly; Microcephaly - genetics; Microcephaly - pathology; Microfilament Proteins - genetics; Microscopy; Mutation; Neurogenesis; NIH 3T3 Cells; Pedigree; Penicillin; Pregnancy; Proteins; Research ethics; Zebrafish; Ciliopathy; CENPF; Clinical genetics; Molecular genetics; Microcephaly
• ISSN: 0022-2593; 1468-6244; 1468-6244
• DOI: 10.1136/jmedgenet-2014-102691

Background Mutations in microtubule-regulating genes are associated with disorders of neuronal migration and microcephaly. Regulation of centriole length has been shown to underlie the pathogenesis of certain ciliopathy phenotypes. Using a next-generation sequencing approach, we identified mutations in a novel centriolar disease gene in a kindred with an embryonic lethal ciliopathy phenotype and in a patient with primary microcephaly. Methods and results Whole exome sequencing data from a non-consanguineous Caucasian kindred exhibiting mid-gestation lethality and ciliopathic malformations revealed two novel non-synonymous variants in CENPF, a microtubule-regulating gene. All four affected fetuses showed segregation for two mutated alleles [IVS5-2A>C, predicted to abolish the consensus splice-acceptor site from exon 6; c.1744G>T, p.E582X]. In a second unrelated patient exhibiting microcephaly, we identified two CENPF mutations [c.1744G>T, p.E582X; c.8692 C>T, p.R2898X] by whole exome sequencing. We found that CENP-F colocalised with Ninein at the subdistal appendages of the mother centriole in mouse inner medullary collecting duct cells. Intraflagellar transport protein-88 (IFT-88) colocalised with CENP-F along the ciliary axonemes of renal epithelial cells in age-matched control human fetuses but did not in truncated cilia of mutant CENPF kidneys. Pairwise co-immunoprecipitation assays of mitotic and serum-starved HEKT293 cells confirmed that IFT88 precipitates with endogenous CENP-F. Conclusions Our data identify CENPF as a new centriolar disease gene implicated in severe human ciliopathy and microcephaly related phenotypes. CENP-F has a novel putative function in ciliogenesis and cortical neurogenesis.