Department of Gynecology, the Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang, China.
Human Reproductive and Genetic Center, Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu, China.
Cell Prolif. 2020 Oct;53(10):e12899. doi: 10.1111/cpr.12899. Epub 2020 Sep 8.
Stem cell niche regulated the renewal and differentiation of germline stem cells (GSCs) in Drosophila. Previously, we and others identified a series of genes encoding ribosomal proteins that may contribute to the self-renewal and differentiation of GSCs. However, the mechanisms that maintain and differentiate GSCs in their niches were not well understood.
Flies were used to generate tissue-specific gene knockdown. Small interfering RNAs were used to knockdown genes in S2 cells. qRT-PCR was used to examine the relative mRNA expression level. TUNEL staining or flow cytometry assays were used to detect cell survival. Immunofluorescence was used to determine protein localization and expression pattern.
Herein, using a genetic manipulation approach, we investigated the role of ribosomal protein S13 (RpS13) in testes and S2 cells. We reported that RpS13 was required for the self-renewal and differentiation of GSCs. We also demonstrated that RpS13 regulated cell proliferation and apoptosis. Mechanistically, we showed that RpS13 regulated the expression of ribosome subunits and could moderate the expression of the Rho1, DE-cad and Arm proteins. Notably, Rho1 imitated the phenotype of RpS13 in both Drosophila testes and S2 cells, and recruited cell adhesions, which was mediated by the DE-cad and Arm proteins.
These findings uncover a novel mechanism of RpS13 that mediates Rho1 signals in the stem cell niche of the Drosophila testis.
干细胞龛调控果蝇生殖干细胞(GSCs)的更新和分化。此前,我们和其他人鉴定了一系列编码核糖体蛋白的基因,这些基因可能有助于 GSCs 的自我更新和分化。然而,维持和分化 GSCs 及其龛位中的机制尚不清楚。
使用果蝇进行组织特异性基因敲低。使用小干扰 RNA(siRNA)在 S2 细胞中敲低基因。qRT-PCR 用于检测相对 mRNA 表达水平。TUNEL 染色或流式细胞术检测细胞存活。免疫荧光用于确定蛋白质定位和表达模式。
在此,我们使用遗传操作方法研究了核糖体蛋白 S13(RpS13)在睾丸和 S2 细胞中的作用。我们报告称,RpS13 是 GSCs 自我更新和分化所必需的。我们还表明,RpS13 调节细胞增殖和凋亡。机制上,我们表明 RpS13 调节核糖体亚基的表达,并能适度调节 Rho1、DE-cad 和 Arm 蛋白的表达。值得注意的是,Rho1 在果蝇睾丸和 S2 细胞中模拟了 RpS13 的表型,并且通过 DE-cad 和 Arm 蛋白募集细胞黏附。
这些发现揭示了 RpS13 在果蝇睾丸干细胞龛中介导 Rho1 信号的新机制。