Long noncoding RNA regulation of spermatogenesis via the spectrin cytoskeleton in Drosophila.
Long noncoding RNA regulation of spermatogenesis via the spectrin cytoskeleton in Drosophila.
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DOI：
10.1093/g3journal/jkab080
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发表时间：
2021-05-07
期刊：
影响因子：
--
通讯作者：
Bai H
中科院分区：
文献类型：
--
作者：
Bouska MJ;Bai H
血影蛋白的细胞骨架已被证明是至关重要的，在不同的过程，如轴突的发育和变性，成肌细胞融合，精子发生。血影蛋白可以通过连接蛋白复合物以组织特异性方式调节，然而，尚未显示长非编码RNA（lncRNA）与血影蛋白相互作用并调节血影蛋白。在这里，我们提供的证据表明，lncRNA CR 45362与α-Spectrin相互作用，是果蝇精子发生过程中精子细胞核捆绑所必需的。我们观察到，CR 45362在睾丸基部的囊肿细胞中显示出高表达，CRISPR介导的CR 45362敲除导致不育雄性，精子细胞核散开，肌动蛋白锥破坏。通过RNA沉淀-质谱（ChIRP-MS）分离染色质，我们鉴定了肌动蛋白-血影蛋白细胞骨架组分与lncRNA CR 45362的物理相互作用。对已鉴定的细胞骨架因子的遗传筛选显示，囊细胞特异性敲低α-Spectrin表型复制CR 45362突变体，并导致精子细胞核束缺陷。结论：CR 45362基因敲除后，α-Spectrin和精子细胞核束在睾丸基底部头部包囊细胞的共定位被破坏。因此，我们发现了一种新的lncRNA CR 45362，它与α-Spectrin相互作用，在精子细胞成熟过程中稳定精子细胞核束。
The spectrin cytoskeleton has been shown to be critical in diverse processes such as axon development and degeneration, myoblast fusion, and spermatogenesis. Spectrin can be modulated in a tissue specific manner through junctional protein complexes, however, it has not been shown that long noncoding RNAs (lncRNAs) interact with and modulate spectrin. Here, we provide evidence of a lncRNA CR45362 that interacts with α-Spectrin, is required for spermatid nuclear bundling during Drosophila spermatogenesis. We observed that CR45362 showed high expression in the cyst cells at the basal testis, and CRISPR-mediated knockout of CR45362 led to sterile male, unbundled spermatid nuclei, and disrupted actin cones. Through chromatin isolation by RNA precipitation—mass spectrometry (ChIRP-MS), we identified actin-spectrin cytoskeletal components physically interact with the lncRNA CR45362. Genetic screening on identified cytoskeletal factors revealed that cyst cell-specific knockdown of α-Spectrin phenocopied CR45362 mutants and resulted in spermatid nuclear bundle defects. Consistently, CR45362 knockout disrupted the co-localization of α-Spectrin and spermatid nuclear bundles in the head cyst cells at the basal testis. Thus, we uncovered a novel lncRNA CR45362 that interacts with α-Spectrin to stabilize spermatid nuclear bundles during spermatid maturation.
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