Mutation of
            <i>serine/threonine protein kinase 36</i>
            (
            <i>STK36</i>
            ) causes primary ciliary dyskinesia with a central pair defect

Edelbusch, Christine; Cindrić, Sandra; Dougherty, Gerard W.; Loges, Niki T.; Olbrich, Heike; Rivlin, Joseph; Wallmeier, Julia; Pennekamp, Petra; Amirav, Israel; Omran, Heymut

doi:10.1002/humu.23261

Cited by 62 publications

(51 citation statements)

References 26 publications

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“…In humans, many axonemal gene mutations of have been shown to cause ciliopathies, such as PCD (Horani and Ferkol, 2018). Historically, conventional TEM analysis of patient ciliary samples is considered a standard clinical diagnostic tool for PCD (Kott et al, 2013;Khouri et al, 2016;Edelbusch et al, 2017). However, the resolution of conventional TEM is greatly limited and could not visualize the defects in the fap216 mutant, hindering the diagnosis of PCD-types caused by minor structural alterations.…”

Section: Fap216 Plays a Role In Chemical Signal Transduction Between mentioning

confidence: 99%

Structural organization of the C1a-e-c supercomplex within the ciliary central apparatus

Zhao

Dymek

et al. 2019

Preprint

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Running title: C1a-e-c supercomplex of motile cilia Summary Fu et al. use a wild-type vs. mutant comparison and cryo-electron tomography of Chlamydomonas flagella to identify central apparatus (CA) subunits and visualize their location in the native 3D CA structure. The study provides a better understanding of the CA and how it regulates ciliary motility. Abbreviations:BCCP, biotin-carboxyl-carrier-protein; CA, central apparatus; CCD, charge-coupled device; cryo-ET, cryo-electron tomography; DMT, doublet microtubule; FAP, flagellar associated protein; IDA, inner dynein arm; MS, mass spectrometry; N-DRC, nexin-dynein regulatory complex; ODA, outer dynein arm; PCD, primary ciliary dyskinesia; psu, peripheral subunit. AbstractNearly all motile cilia contain a central apparatus (CA) composed of two connected singletmicrotubules with attached projections that play crucial roles in regulating ciliary motility.Defects in CA assembly usually result in motility-impaired or paralyzed cilia, which in humans causes disease. Despite their importance, the protein composition and functions of the CA projections are largely unknown. Here, we integrated biochemical and genetic approaches with cryo-electron tomography to compare the CA of wild type Chlamydomonas with CA mutants.We identified a large (>2 MDa) complex, the C1a-e-c supercomplex, that requires the PF16 protein for assembly and contains the CA components FAP76, FAP81, FAP92, and FAP216. We localized these subunits within the supercomplex using nanogold-labeling and show that loss of any one of them results in impaired ciliary motility. These data provide insight into the subunit organization and three-dimensional (3D) structure of the CA, which is a prerequisite for understanding the molecular mechanisms by which the CA regulates ciliary beating. Despite many biochemical and structural studies of the CA (Witman et al., 1978; protein composition, 3D organization, and functional mechanism(s) of the CA in ciliary motility are not fully understood. Our recent mass spectrometry (MS) study compared the proteomes of Chlamydomonas wild-type and mutant axonemes, and identified 44 new candidate CA proteins assigned to the C1 or the C2 microtubule (Zhao et al., 2019).However, questions about the organization, assembly, and function of the CA and its projections remain, making the CA the structurally and functionally least understood axonemal complex to date.Here we combined biochemical, genetic, and structural analyses to investigate the protein composition and molecular organization of a group of interconnected CA projections, here termed the C1a-e-c supercomplex, in wild-type and CA mutants of Chlamydomonas. Sucrose gradient sedimentation and MS revealed that several CA proteins identified in this study, FAP76, FAP81, FAP92 and FAP216, are associated with the protein PF16, previously assigned to the C1 microtubule but not to a specific projection. Chlamydomonas mutants that lacked any of these proteins showed impaired motility. Structural comparisons of flagella from wild-ty...

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Section: Fap216 Plays a Role In Chemical Signal Transduction Between mentioning

confidence: 99%

Structural organization of the C1a-e-c supercomplex within the ciliary central apparatus

Zhao

Dymek

et al. 2019

Preprint

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“…The Pcdo allele recapitulated most PCD phenotypes previously observed in other models due to central pair abnormalities. These are often the most difficult to diagnose in human due to lack of laterality phenotypes, only subtle beating defects and largely undisturbed cilia ultrastructure (Edelbusch, Cindrić et al 2017). We suggest Spag17 Pcdo is very useful model to study the pathogenesis and molecular mechanisms employed in the development of human PCD subsequent to central pair defects.…”

Section: Ependymal Cilia Show Normal Ultrastructure and Minimal Beatimentioning

confidence: 98%

A novel hypomorphic allele ofSpag17causes primary ciliary dyskinesia phenotypes in mice

Abdelhamed

Lukacs

Cindrić

et al. 2020

Preprint

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Primary ciliary dyskinesia (PCD) is a human condition of dysfunctional motile cilia characterized by recurrent lung infection, infertility, organ laterality defects, and partially penetrant hydrocephalus. We recovered a mouse mutant from a forward genetic screen that developed all the phenotypes of PCD. Whole exome sequencing identified this primary ciliary dyskinesia only (Pcdo) allele to be a nonsense mutation (c.5236A>T) in the Spag17 coding sequence creating a premature stop codon at position 1746 (K1746*). The Pcdo variant abolished different isoforms of SPAG17 in the Pcdo mutant testis but not in the brain. Our data indicate differential requirements for SPAG17 in different motile cilia cell types. SPAG17 is required for proper development of the sperm flagellum, and is essential for either development or stability of the C1 microtubule structure within cilia, but not the brain ependymal cilia. We identified changes in ependymal cilia beating frequency but these did not apparently alter lateral ventricle cerebrospinal fluid (CSF) flow. Aqueductal (Aq) stenosis resulted in significantly slower and abnormally directed CSF flow and we suggest this is the root cause of the hydrocephalus. The Spag17Pcdo homozygous mutant mice are generally viable to adulthood, but have a significantly shortened life span with chronic morbidity. Our data indicate that the c.5236A>T Pcdo variant is a hypomorphic allele of Spag17 gene that causes phenotypes related to motile, but not primary, cilia. Spag17Pcdo is a novel and useful model for elucidating the molecular mechanisms underlying development of PCD in the mouse.

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“…In addition, mutations in two genes, CCNO (15) and MCIDAS (16), cause a PCD-like phenotype by greatly reducing the number of motile cilia. Although genetic variants that cause PCD have been identified in over 40 genes (2,(17)(18)(19)(20)(21)(22)(23), there are individuals with confirmed clinical features of PCD but normal axonemal structure as determined by transmission electron microscopy, for whom the genetic basis of their disease is unknown.…”

Section: Introductionmentioning

confidence: 99%

Mutation ofCFAP57causes primary ciliary dyskinesia by disrupting the asymmetric targeting of a subset of ciliary inner dynein arms

Bustamante-Marin

Horani

Stoyanova

et al. 2019

Preprint

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word count: 247 Text word count, without methods: 3856 Abstract (247/ 250 words)Primary ciliary dyskinesia (PCD) is characterized by chronic airway disease, male infertility, and randomization of the left/right body axis, and is caused by defects of motile cilia and sperm flagella. We screened a cohort of affected individuals that lack an obvious TEM structural phenotype for pathogenic variants using whole exome capture and next generation sequencing. The population sampling probability (PSAP) algorithm identified one subject with a homozygous nonsense variant [(c.1762C>T) p.(Arg588*) exon 11] in the uncharacterized CFAP57 gene. In normal human nasal epithelial cells, CFAP57 localizes throughout the ciliary axoneme. Analysis of cells from the PCD patient shows a loss of CFAP57, reduced beat frequency, and an alteration in the ciliary waveform. Knockdown of CFAP57 in human tracheobronchial epithelial cells (hTECs)recapitulates these findings. Phylogenetic analysis showed that CFAP57 is conserved in organisms that assemble motile cilia, and CFAP57 is allelic with the BOP2 gene identified previously in Chlamydomonas. Two independent, insertional fap57Chlamydomonas mutant strains show reduced swimming velocity and altered waveforms. Tandem mass spectroscopy showed that CFAP57 is missing, and the "g" inner dyneins (DHC7 and DHC3) and the "d" inner dynein (DHC2) are reduced. Our data demonstrate that the FAP57 protein is required for the asymmetric assembly of inner dyneins on only a subset of the microtubule doublets, and this asymmetry is essential for the generation of an effective axonemal waveform. Together, our data identifies mutations in CFAP57 as a cause of PCD with a specific defect in the inner dynein arm assembly process. Significance (119/120 words)Motile cilia are found throughout eukaryotic organisms and performs essential functions.Primary ciliary dyskinesia (PCD) is a rare disease that affects the function of motile cilia.By applying a novel population sampling probability algorithm (PSAP) that uses large population sequencing databases and pathogenicity prediction algorithms, we identified a variant in an uncharacterized gene, CFAP57. This is the first reported example of PCD caused by a mutation that affects only a subset of the inner dynein arms, which are needed to generate the waveform. CFAP57 identifies an address for specific dynein arms. These findings demonstrate the effectiveness of the PSAP algorithm, expand our understanding of the positioning of dynein arms, and identify mutations in CFAP57 as a cause of PCD.

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Mutation of serine/threonine protein kinase 36 ( STK36 ) causes primary ciliary dyskinesia with a central pair defect

Cited by 62 publications

References 26 publications

Structural organization of the C1a-e-c supercomplex within the ciliary central apparatus

Structural organization of the C1a-e-c supercomplex within the ciliary central apparatus

A novel hypomorphic allele ofSpag17causes primary ciliary dyskinesia phenotypes in mice

Mutation ofCFAP57causes primary ciliary dyskinesia by disrupting the asymmetric targeting of a subset of ciliary inner dynein arms

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