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

Abstract

BackgroundMutations 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 resultsWhole 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.ConclusionsOur 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.

Strømme Syndrome Is a Ciliary Disorder Caused by Mutations inCENPF

“…Compound heterozygous inheritance was confirmed in both families. Recently, mutations in this gene were shown to cause a fetal lethal phenotype, the phenotype and functional data being compatible with a human ciliopathy [Waters et al., ]. We show for the first time that Strømme syndrome is an autosomal‐recessive disease caused by mutations in CENPF that can result in a wide phenotypic spectrum.…”

“…()) represents the severe end of the phenotypic spectrum. An intermediate form allows prolonged survival of individuals after surgical repair of intestinal atresias (Family A in this study), whereas nonsyndromic microcephaly seems to represent a milder ciliopathy phenotype [Waters et al., ]. On the contrary, embryonic arrest at the 8‐cell stage in bovine preimplantation embryos due to depletion of CENPF with a dramatic decrease of developmental competence after embryonic genome activation [Toralová et al., ] may be considered to be an extreme lethal phenotype.…”

Centriolar satellites are acentriolar assemblies of centrosomal proteins

“…For most proteins, the trend reflected results from the WCP (i.e. PLK4 is up‐, whereas CEP63 and CEP57 down‐regulated) with the ciliopathy‐associated proteins CEP41 and CENPF being notable exceptions (Lee et al , ; Waters et al , ). Despite no change in overall protein levels, CEP41 was 16‐fold down‐regulated in SILAC‐CS‐STIL, whereas CENPF exhibited a 2‐fold increase in SILAC‐CS‐STIL (Fig E–G).…”

CPAP promotes timely cilium disassembly to maintain neural progenitor pool

“…Recent studies have uncovered that the proper timing of cilium disassembly mediated by Nde‐1 and TcTex‐1 is critically regulated during cell cycle (Kim et al , 2011; Li et al , 2011; Maskey et al , 2015). Interestingly, mutations in Nde‐1 and microtubule and kinetochore components causing microcephaly and ciliary defects highlight a possible role for the cilium in regulating NPCs to control the number of neurons generated during brain development (Alkuraya et al , 2011; Hu et al , 2014; Waters et al , 2015). However, the biological significance of these spatiotemporally interlinked processes namely cilium disassembly, cell cycle re‐entry and neural progenitor cells (NPCs) differentiation, and the molecular mechanisms linking these cellular events together remain unclear.…”