CEP164-null cells generated by genome editing show a ciliation defect with intact DNA repair capacity
CEP164-null cells generated by genome editing show a ciliation defect with intact DNA repair capacity
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DOI：
10.1242/jcs.186221
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发表时间：
2016-05-01
影响因子：
4
通讯作者：
Morrison, Ciaran G.
中科院分区：
文献类型：
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作者：
Daly, Owen M.;Gaboriau, David;Morrison, Ciaran G.
初级纤毛是从成熟的中心粒末端延伸出来的微管结构。CEP164是一个由母中心粒携带的远端附属物的组成部分，是初级纤毛形成所必需的。最近的数据表明CEP164是一个纤毛病变基因，并表明CEP164在DNA损伤反应（DDR）中起着一定的作用。我们使用反向遗传学来测试CEP164在DDR中的作用。我们发现，使用生长素诱导的降解决定子在鸡DT 40细胞中有条件地消耗CEP164导致对电离或紫外线照射诱导的DNA损伤的敏感性没有增加。CEP164在人视网膜色素上皮细胞中的破坏阻断了初级纤毛的形成，但不影响细胞增殖或细胞对电离或紫外线照射的反应。此外，我们使用免疫荧光显微镜和多种标记形式的CEP164的分析没有观察到CEP164定位到细胞核。我们的数据表明，CEP164在DDR中不需要。
Primary cilia are microtubule structures that extend from the distal end of the mature, mother centriole. CEP164 is a component of the distal appendages carried by the mother centriole that is required for primary cilium formation. Recent data have implicated CEP164 as a ciliopathy gene and suggest that CEP164 plays some roles in the DNA damage response (DDR). We used reverse genetics to test the role of CEP164 in the DDR. We found that conditional depletion of CEP164 in chicken DT40 cells using an auxin-inducible degron led to no increase in sensitivity to DNA damage induced by ionising or ultraviolet irradiation. Disruption of CEP164 in human retinal pigmented epithelial cells blocked primary cilium formation but did not affect cellular proliferation or cellular responses to ionising or ultraviolet irradiation. Furthermore, we observed no localisation of CEP164 to the nucleus using immunofluorescence microscopy and analysis of multiple tagged forms of CEP164. Our data suggest that CEP164 is not required in the DDR.