BmGeminin2 interacts with BmRRS1 and regulates Bombyx mori cell proliferation
BmGeminin2 interacts with BmRRS1 and regulates Bombyx mori cell proliferation
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BmGeminin2 与 BmRRS1 相互作用并调节家蚕细胞增殖
DOI：
10.1080/15384101.2019.1624109
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发表时间：
2019
期刊：
影响因子：
4.3
通讯作者：
Lu Cheng
中科院分区：
文献类型：
--
作者：
Zhou Xiao Lin;Wei Yi;Chen Xiang Yun;Chen Peng;Tang Xiao Fang;Zhang Qian;Dong Zhan Qi;Pan Min Hui;Lu Cheng
摘要在脊椎动物和无脊椎动物中，双子蛋白是细胞周期进程的主要调节因子，它确保DNA复制的及时启动并防止再复制。在此之前，我们在家蚕中鉴定了两个Geminin基因，BmGeminin 1和BmGemin 2，并且我们发现BmGemin 1的RNA干扰导致了重新复制。然而，BmGeminin 2的功能仍然知之甚少。本研究发现BmGeminin 2基因敲低可促进细胞增殖，并上调G2/M期相关基因cyclinB/CDK 1的表达。然后，我们进行酵母双杂交筛选，以确定相互作用的蛋白质。我们的研究结果产生了23个相互作用的蛋白质，它们涉及DNA复制、染色体稳定、胚胎发育、能量、防御、蛋白质加工或结构蛋白。在这里，我们专注于BmRRS 1，这是一种与细胞周期G2/M进程密切相关的染色体大会相关蛋白。通过免疫荧光和免疫沉淀证实了BmGeminin 2和BmRRS 1之间的相互作用。其表达谱分析显示BmRRS 1与BmGeminin 2相关。此外，BmGeminin 2过表达下调BmRRS 1转录。BmGeminin 2的敲低导致BmRRS 1转录物的上调。此外，BmRRS 1的过表达可上调G2/M期相关基因cyclinB/CDK 1的表达，促进细胞增殖，与BmGeminin 2基因敲除的作用一致。此外，BmRRS 1 RNA干扰可以消除BmGem 2基因敲除对细胞增殖、细胞周期分期比例和cyclinB/CDK 1表达的影响。这些数据表明，BmGeminin 2基因敲除的细胞增殖优势与BmRRS 1密切相关。我们的研究结果提供了深入了解Geminin的功能和调控家蚕细胞周期的机制。
ABSTRACT Geminin is a master regulator of cell-cycle progression that ensures the timely onset of DNA replication and prevents re-replication in vertebrates and invertebrates. Previously, we identified two Geminin genes, BmGeminin1 and BmGeminn2, in the silkworm Bombyx mori, and we found that RNA interference of BmGeminin1 led to re-replication. However, the function of BmGeminin2 remains poorly understood. In this study, we found that knockdown of BmGeminin2 can improve cell proliferation, and upregulated G2/M-associated gene-cyclinB/CDK1 expression. Then, we performed yeast two-hybrid screening to identify interacting proteins. Our results yielded 23 interacting proteins, which are involved in DNA replication, chromosome stabilization, embryonic development, energy, defense, protein processing, or structural protein. Here, we focused on BmRRS1, a chromosome congression-related protein that is closely related to cell cycle G2/M progression. The interaction between BmGeminin2 and BmRRS1 was confirmed by immunofluorescence and immunoprecipitation. Analysis of its expression profile showed that BmRRS1 was related to BmGeminin2. In addition, BmGeminin2 overexpression downregulated the BmRRS1 transcript. Knockdown of BmGeminin2 led to upregulation of the BmRRS1 transcript. Furthermore, overexpression of BmRRS1 can upregulate G2/M-associated gene-cyclinB/CDK1 expression, and improved cell proliferation, consistent with the effects of BmGeminin2 knockout. In addition, BmRRS1 RNA interference can eliminate the impact of BmGem2 knockout on cell proliferation, the ratio of cell cycle stage and the expression of cyclinB/CDK1. These data suggested that the cell proliferation advantage of BmGeminin2 knockout was closely related to BmRRS1. Our findings provide insight into the functions of Geminin and the mechanisms underlying the regulation of the cell cycle in the silkworm.