The SON RNA splicing factor is required for intracellular trafficking structures that promote centriole assembly and ciliogenesis,Molecular Biology of the Cell

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The SON RNA splicing factor is required for intracellular trafficking structures that promote centriole assembly and ciliogenesis
Molecular Biology of the Cell ( IF 3.1 ) Pub Date : 2021-08-18 , DOI: 10.1091/mbc.e21-06-0305
Alexander J Stemm-Wolf ₁ , Eileen T O'Toole ₂ , Ryan M Sheridan ₃ , Jacob T Morgan ₁ , Chad G Pearson ₁

Affiliation

Control of centrosome assembly is critical for cell division, intracellular trafficking and cilia. Regulation of centrosome number occurs through the precise duplication of centrioles that reside in centrosomes. Here we explored transcriptional control of centriole assembly and find that the RNA splicing factor SON is specifically required for completing procentriole assembly. Whole genome mRNA sequencing identified genes whose splicing and expression are affected by the reduction of SON, with an enrichment in genes involved in the microtubule cytoskeleton, centrosome and centriolar satellites. SON is required for the proper splicing and expression of CEP131 which encodes a major centriolar satellite protein and is required to organize the trafficking and microtubule network around the centrosomes. This study highlights the importance of the distinct microtubule trafficking network that is intimately associated with nascent centrioles and is responsible for procentriole development and efficient ciliogenesis.

Movie S1: Serial tomographic slices through a volume containing an siControl rosette. The tomographic volume as built from three, serial 250 thick sections. This centrosome is displayed in Figure 3B and Figure Supplement 7A (A′). The centrosome is close to the nucleus and numerous nuclear pore complexes can be observed in the serial tomographic slices. The model shows two mother centrioles (green), and four procentrioles (blue). Microtubules (magenta) and microtubule minus ends (yellow) are modelled. Bar = 200 nm.Download Original Video (8.0 MB)

Movie S2: Serial tomographic slices through a volume containing the centrosome of an siSON cell. A tomogram from an siSON cell shown in Figure 3B and Figure Supplement 7A (D″). The tomographic volume as built from four, serial 250 thick sections. Vesicles are present in this region around the two mother centrioles (green) and six procentrioles (blue) present in the volume. Microtubules (magenta) and microtubule minus ends (yellow) are modelled. Bar = 200 nm.Download Original Video (6.0 MB)

Movie S3: Serial, tomographic slices through a volume containing the centrosome of an siControl cell. A partial reconstruction through a volume containing the centrosome of the siControl cell shown in Figure 7E and Figure Supplement 7A (B′). This volume was built from one, 250nm thick section and therefore the mother centriole (green) and five procentrioles (blue) are not complete in the volume. Microtubules (magenta) and microtubule minus ends (yellow) are modelled. Bar = 200 nm.Download Original Video (4.9 MB)

Movie S4: Serial tomographic slices through a volume containing one centrosome of an siSON cell. This centrosome is shown in Figure 7E and Figure Supplement 7A (A″ left). The tomographic volume as built from two, serial 250 thick sections. The model shows 1 mature centriole (green) with distal and subdistal appendages present. Microtubules can be seen originating at appendages (modelled in magenta with minus ends shown in yellow). Bar = 200 nm.Download Original Video (5.6 MB)

Movie S5: Serial tomographic slices through a volume containing one centrosome of an siSON cell. This is the second centrosome from the same cell depicted in Movie S4 and is shown in Figure Supplement 7A (A″ right). The tomographic volume as built from two, serial 250 thick sections. No detectable procentrioles were observed, although a mass of electron dense material was observed adjacent to one of the mother centrioles. The centriole is modelled in green, the microtubules in magenta and the microtubule minus ends in yellow. Bar = 200 nm.Download Original Video (5.8 MB)

Movie S6: Serial tomographic slices through a volume containing two centrosomes of an siSON cell. The tomographic volume as built from two, serial 250 thick sections and shown in Figure Supplement 7A (B″). Two mature centrioles (green) are observed, one of which has distal and subdistal appendages. Vesicles are present throughout the volume. The volume does not contain the entire length of one of these centrioles. Each mature centriole has a procentriole associated with it (blue). Microtubules (magenta) and microtubule minus ends (yellow) are also modelled. Bar = 200 nm.Download Original Video (5.8 MB)

Movie S7: Serial tomographic slices through a volume containing two centrosomes of an siSON cell. The tomographic volume as built from four, serial 250 thick sections shown in Figure Supplement 7A (C″). Two mature centrioles (green) are observed, one of which has distal and subdistal appendages. Vesicles are present throughout the volume. Each mature centriole has a procentriole associated with it (blue). Microtubules (magenta) and microtubule minus ends (yellow) are also modelled. Bar = 200 nm.Download Original Video (5.7 MB)

Movie S8: Serial tomographic slices through a volume containing three elongated centrioles in an siControl cell. The tomographic volume as built from four, serial 250 thick sections and displayed in Figure Supplement 7A (C′). Three fully elongated centrioles are observed (green). Microtubules (magenta) and microtubule minus ends (yellow) are also modelled. Bar = 200 nm.Download Original Video (8.3 MB)

Movie S9: Serial tomographic slices through a volume containing one of two centrosomes in an siControl cell. The tomographic volume as built from three, serial 250 thick sections and displayed in Figure Supplement 7A (D′). The second centrosome can be seen in Movie S10. One mature centriole (green) and three procentrioles (blue) are present in the volume. Microtubules (magenta) and microtubule minus ends (yellow) are also modelled. Bar = 200 nm.Download Original Video (5.1 MB)

Movie S10: Serial tomographic slices through a volume containing the second of two centrosomes in an siControl cell. The tomographic volume as built from two, serial 250 thick sections and displayed in Figure Supplement 7A (E′). One mature centriole (green) and two procentrioles (blue) are present in the volume. The other centrosome from this cell is shown in Movie S9. Microtubules (magenta) and microtubule minus ends (yellow) are also modelled. Bar = 200 nm.Download Original Video (3.5 MB)

中文翻译：

SON RNA 剪接因子是促进中心粒组装和纤毛发生的细胞内运输结构所必需的

中心体组装的控制对于细胞分裂、细胞内运输和纤毛至关重要。中心体数量的调节是通过中心体中中心粒的精确复制来实现的。在这里，我们探索了中心粒组装的转录控制，发现RNA剪接因子SON是完成原中心粒组装所特别需要的。全基因组 mRNA 测序鉴定了剪接和表达受 SON 减少影响的基因，其中涉及微管细胞骨架、中心体和中心粒卫星的基因富集。SON 是 CEP131 正确剪接和表达所必需的，CEP131编码主要的中心粒卫星蛋白，并且是组织中心体周围的运输和微管网络所必需的。这项研究强调了独特的微管运输网络的重要性，该网络与新生中心粒密切相关，并负责原中心粒的发育和有效的纤毛发生。

影片 S1：通过包含 siControl 玫瑰花结的体积的串行断层扫描切片。断层扫描体由三个连续的 250 个厚截面组成。该中心体显示在图 3B 和图补充 7A (A') 中。中心体靠近细胞核，在连续断层切片中可以观察到许多核孔复合体。该模型显示两个母中心粒（绿色）和四个原中心粒（蓝色）。微管（洋红色）和微管负端（黄色）已建模。Bar = 200 nm。下载原始视频 (8.0 MB)

影片 S2：通过包含 siSON 细胞中心体的体积的连续断层扫描切片。图 3B 和图补充 7A (D") 中所示的 siSON 电池断层图。断层扫描体积由四个连续的 250 个厚切片组成。囊泡存在于体积中存在的两个母中心粒（绿色）和六个原中心粒（蓝色）周围的区域中。微管（洋红色）和微管负端（黄色）已建模。Bar = 200 nm。下载原始视频 (6.0 MB)

影片 S3：通过包含 siControl 细胞中心体的体积进行连续断层扫描切片。通过包含 siControl 细胞中心体的体积进行部分重建，如图 7E 和图补充 7A (B') 所示。该体积是由一个 250 nm 厚的切片构建的，因此该体积中的母中心粒（绿色）和五个原中心粒（蓝色）并不完整。微管（洋红色）和微管负端（黄色）已建模。Bar = 200 nm。下载原始视频 (4.9 MB)

影片 S4：通过含有 siSON 细胞中心体的体积的连续断层扫描切片。该中心体如图 7E 和补充图 7A 所示（左 A”）。断层扫描体由两个连续的 250 个厚截面构成。该模型显示 1 个成熟的中心粒（绿色），具有远端和近远端附属物。可以看到微管起源于附属物（以洋红色建模，负端以黄色显示）。Bar = 200 nm。下载原始视频 (5.6 MB)

影片 S5：通过含有 siSON 细胞中心体的体积的连续断层扫描切片。这是来自电影 S4 中描绘的同一细胞的第二个中心体，如图补充 7A（A“右）所示。断层扫描体由两个连续的 250 个厚截面构成。尽管在母中心粒之一附近观察到大量电子致密材料，但没有观察到可检测到的原中心粒。中心粒以绿色建模，微管以洋红色建模，微管负端以黄色建模。Bar = 200 nm。下载原始视频 (5.8 MB)

影片 S6：通过含有 siSON 细胞两个中心体的体积的连续断层扫描切片。断层扫描体积由两个连续的 250 个厚截面构成，如图补充 7A (B") 所示。观察到两个成熟的中心粒（绿色），其中之一具有远端和近远端附属物。囊泡存在于整个体积中。该体积不包含这些中心粒之一的整个长度。每个成熟的中心粒都有一个与之相关的原中心粒（蓝色）。还对微管（洋红色）和微管负端（黄色）进行了建模。Bar = 200 nm。下载原始视频 (5.8 MB)

影片 S7：通过包含 siSON 细胞两个中心体的体积的连续断层扫描切片。断层扫描体积由四个连续的 250 个厚截面构成，如图补充 7A (C") 所示。观察到两个成熟的中心粒（绿色），其中之一具有远端和近远端附属物。囊泡存在于整个体积中。每个成熟的中心粒都有一个与之相关的原中心粒（蓝色）。还对微管（洋红色）和微管负端（黄色）进行了建模。Bar = 200 nm。下载原始视频 (5.7 MB)

影片 S8：连续断层扫描切片穿过 siControl 单元中包含三个细长中心粒的体积。断层扫描体积由四个连续的 250 个厚切片组成，并显示在图补充 7A (C') 中。观察到三个完全拉长的中心粒（绿色）。还对微管（洋红色）和微管负端（黄色）进行了建模。Bar = 200 nm。下载原始视频 (8.3 MB)

影片 S9：通过包含 siControl 细胞中两个中心体之一的体积的连续断层扫描切片。断层扫描体积由三个连续的 250 个厚切片组成，并显示在图补充 7A (D') 中。第二个中心体可以在电影S10中看到。体积中存在一个成熟的中心粒（绿色）和三个原中心粒（蓝色）。还对微管（洋红色）和微管负端（黄色）进行了建模。Bar = 200 nm。下载原始视频 (5.1 MB)

影片 S10：通过包含 siControl 细胞中两个中心体中的第二个的体积的连续断层扫描切片。断层扫描体积由两个连续的 250 个厚切片组成，并显示在图补充 7A (E') 中。体积中存在一个成熟的中心粒（绿色）和两个原中心粒（蓝色）。电影 S9 中显示了该细胞的另一个中心体。还对微管（洋红色）和微管负端（黄色）进行了建模。Bar = 200 nm。下载原始视频 (3.5 MB)

更新日期：2021-08-19

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