Functions and substrates of NEDDylation during cell cycle in the silkworm, Bombyx mori.
Functions and substrates of NEDDylation during cell cycle in the silkworm, Bombyx mori.
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DOI：
10.1016/j.ibmb.2017.09.013
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发表时间：
2017-11
影响因子：
3.8
通讯作者：
Zhiqing Li;Qixin Cui;Xiao-yan Wang;Bingqian Li;Dongchao Zhao;Q. Xia;P. Zhao
中科院分区：
文献类型：
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作者：
Zhiqing Li;Qixin Cui;Xiao-yan Wang;Bingqian Li;Dongchao Zhao;Q. Xia;P. Zhao
NEDD化是由泛素样蛋白Nedd 8与特定底物结合介导的翻译后修饰，是调节真核生物细胞周期进程的重要生物学过程。在这里，我们报告的NEDD化机制和NEDDylated蛋白质在家蚕，家蚕的保护。我们已经鉴定了家蚕中可逆NEDD化所需的所有组分，包括Nedd 8、E1、E2、E3和deNEDD化酶。通过RNAi介导的基因沉默方法，表明BmNedd 8和缀合酶的敲低降低了培养的家蚕细胞中NEDD化的总体水平，而deNEDD化酶的敲低增加了这种修饰的流行。此外，NEDD化系统的缺乏导致细胞周期停滞在G2/M期，并导致染色体聚集和分离的缺陷。利用BmNedd 8的野生型和突变体，我们鉴定了BmNedd 8的特异性底物，其参与许多细胞过程的调节，包括核糖体生物合成、剪接体结构、纺锤体形成、代谢和RNA生物合成。这清楚地表明，NEDD化系统能够控制家蚕中的多种途径。总之，本文所提供的关于NEDD化系统的功能和底物的信息可以为将来研究家蚕蛋白质NEDD化及其对细胞周期进程的调控机制提供基础。
NEDDylation, a post-translational modification mediated by the conjugation of the ubiquitin-like protein Nedd8 to specific substrates, is an essential biological process that regulates cell cycle progression in eukaryotes. Here, we report the conservation of NEDDylation machinery and NEDDylated proteins in the silkworm,Bombyx mori. We have identified all the components necessary for reversible NEDDylation in the silkworm including Nedd8, E1, E2, E3, and deNEDDylation enzymes. By the approach of RNAi-mediated gene silencing, it was shown that knockdown of BmNedd8 and the conjugating enzymes decreased the global level of NEDDylation, while knockdown of deNEDDylation enzymes increased the prevalence of this modification in cultured silkworm cells. Moreover, the lack of the NEDDylation system caused cell cycle arrest at the G2/M phase and resulted in defects in chromosome congression and segregation. Using the wild-type and mutants of BmNedd8, we identified the specific substrates of BmNedd8, which are involved in the regulation for many cellular processes, including ribosome biogenesis, spliceosome structure, spindle formation, metabolism, and RNA biogenesis. This clearly demonstrates that the NEDDylation system is able to control multiple pathways in the silkworm. Altogether, the information on the functions and substrates of the NEDDylation system presented here could provide a basis for future investigations of protein NEDDylation and its regulatory mechanism on cell cycle progression in the silkworm.