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      Ciliary transcription factors and miRNAs precisely regulate Cp110 levels required for ciliary adhesions and ciliogenesis

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          Abstract

          Upon cell cycle exit, centriole-to-basal body transition facilitates cilia formation. The centriolar protein Cp110 is a regulator of this process and cilia inhibitor, but its positive roles in ciliogenesis remain poorly understood. Using Xenopus we show that Cp110 inhibits cilia formation at high levels, while optimal levels promote ciliogenesis. Cp110 localizes to cilia-forming basal bodies and rootlets, and is required for ciliary adhesion complexes that facilitate Actin interactions. The opposing roles of Cp110 in ciliation are generated in part by coiled-coil domains that mediate preferential binding to centrioles over rootlets. Because of its dual role in ciliogenesis, Cp110 levels must be precisely controlled. In multiciliated cells, this is achieved by both transcriptional and post-transcriptional regulation through ciliary transcription factors and microRNAs, which activate and repress cp110 to produce optimal Cp110 levels during ciliogenesis. Our data provide novel insights into how Cp110 and its regulation contribute to development and cell function.

          DOI: http://dx.doi.org/10.7554/eLife.17557.001

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          Most cited references44

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          The genome of the Western clawed frog Xenopus tropicalis.

          The western clawed frog Xenopus tropicalis is an important model for vertebrate development that combines experimental advantages of the African clawed frog Xenopus laevis with more tractable genetics. Here we present a draft genome sequence assembly of X. tropicalis. This genome encodes more than 20,000 protein-coding genes, including orthologs of at least 1700 human disease genes. Over 1 million expressed sequence tags validated the annotation. More than one-third of the genome consists of transposable elements, with unusually prevalent DNA transposons. Like that of other tetrapods, the genome of X. tropicalis contains gene deserts enriched for conserved noncoding elements. The genome exhibits substantial shared synteny with human and chicken over major parts of large chromosomes, broken by lineage-specific chromosome fusions and fissions, mainly in the mammalian lineage.
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            In situ hybridization: an improved whole-mount method for Xenopus embryos.

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              Cep97 and CP110 suppress a cilia assembly program.

              Mammalian centrioles play a dynamic role in centrosome function, but they also have the capacity to nucleate the assembly of cilia. Although controls must exist to specify these different fates, the key regulators remain largely undefined. We have purified complexes associated with CP110, a protein that plays an essential role in centrosome duplication and cytokinesis, and have identified a previously uncharacterized protein, Cep97, that recruits CP110 to centrosomes. Depletion of Cep97 or expression of dominant-negative mutants results in CP110 disappearance from centrosomes, spindle defects, and polyploidy. Remarkably, loss of Cep97 or CP110 promotes primary cilia formation in growing cells, and enforced expression of CP110 in quiescent cells suppresses their ability to assemble cilia, suggesting that Cep97 and CP110 collaborate to inhibit a ciliogenesis program. Identification of Cep97 and other genes involved in regulation of cilia assembly may accelerate our understanding of human ciliary diseases, including renal disease and retinal degeneration.
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                Author and article information

                Contributors
                Role: Reviewing editor
                Journal
                eLife
                Elife
                eLife
                eLife
                eLife
                eLife Sciences Publications, Ltd
                2050-084X
                13 September 2016
                2016
                : 5
                : e17557
                Affiliations
                [1 ]deptDivision of Genetics, Genomics and Development, Center for Integrative Genomics, Department of Molecular and Cell Biology , University of California , Berkeley, United States
                [2 ]deptMolecular Neurobiology Laboratory , Salk Institute for Biological Studies , La Jolla, United States
                [3]University of Toronto , Canada
                Author notes
                Author information
                http://orcid.org/0000-0002-2332-6068
                http://orcid.org/0000-0003-0075-8324
                http://orcid.org/0000-0001-8247-4880
                Article
                17557
                10.7554/eLife.17557
                5045295
                27623009
                113833d3-eb70-444d-b70b-6c7397c1f94a
                © 2016, Walentek et al

                This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

                History
                : 05 May 2016
                : 12 September 2016
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001659, Deutsche Forschungsgemeinschaft;
                Award ID: Wa 3365/1-1
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100000050, National Heart, Lung, and Blood Institute;
                Award ID: K99HL127275
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100000057, National Institute of General Medical Sciences;
                Award ID: GM42341
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100000057, National Institute of General Medical Sciences;
                Award ID: GM076507
                Award Recipient :
                The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
                Categories
                Cell Biology
                Developmental Biology and Stem Cells
                Research Article
                Custom metadata
                2.5
                At optimal concentrations, the ciliary inhibitor Cp110 promotes ciliogenesis by localization to previously uncharacterized sites at the basal body, where it recruits ciliary adhesion complexes that mediate basal body interaction with F-actin networks.

                Life sciences
                centriole,mucociliary epithelia,mir-34,mir-449,foxj1,rfx2,mcidas,focal adhesion,cilia,xenopus,human
                Life sciences
                centriole, mucociliary epithelia, mir-34, mir-449, foxj1, rfx2, mcidas, focal adhesion, cilia, xenopus, human

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