Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg 41390, Sweden.
Proc Natl Acad Sci U S A. 2022 Oct 11;119(41):e2207605119. doi: 10.1073/pnas.2207605119. Epub 2022 Oct 3.
The cilium-centrosome complex contains triplet, doublet, and singlet microtubules. The lumenal surfaces of each microtubule within this diverse array are decorated by microtubule inner proteins (MIPs). Here, we used single-particle cryo-electron microscopy methods to build atomic models of two types of human ciliary microtubule: the doublet microtubules of multiciliated respiratory cells and the distal singlet microtubules of monoflagellated human spermatozoa. We discover that SPACA9 is a polyspecific MIP capable of binding both microtubule types. SPACA9 forms intralumenal striations in the B tubule of respiratory doublet microtubules and noncontinuous spirals in sperm singlet microtubules. By acquiring new and reanalyzing previous cryo-electron tomography data, we show that SPACA9-like intralumenal striations are common features of different microtubule types in animal cilia. Our structures provide detailed references to help rationalize ciliopathy-causing mutations and position cryo-EM as a tool for the analysis of samples obtained directly from ciliopathy patients.
纤毛-中心体复合物包含三联体、二联体和单体微管。在这个多样化的阵列中,每个微管的内腔表面都由微管内蛋白(MIP)修饰。在这里,我们使用单颗粒冷冻电子显微镜方法构建了两种类型的人纤毛微管的原子模型:多纤毛呼吸细胞的二联体微管和单鞭毛人类精子的远端单体微管。我们发现 SPACA9 是一种多特异性 MIP,能够结合两种微管类型。SPACA9 在呼吸二联体微管的 B 管腔内形成内腔条纹,并在精子单体微管中形成非连续螺旋。通过获取新的和重新分析以前的冷冻电子断层扫描数据,我们表明 SPACA9 样内腔条纹是动物纤毛中不同微管类型的共同特征。我们的结构提供了详细的参考,有助于合理化纤毛病突变,并将冷冻电镜定位为直接从纤毛病患者获得的样本的分析工具。
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