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RNA干扰介导的基因mN6A1敲低可降低细胞增殖并减少蛋白质翻译。

RNAi-mediated knock-down of gene mN6A1 reduces cell proliferation and decreases protein translation.

作者信息

Liu Yongbo, Nie Dongsong, Huang Yuelong, Lu Guangxiu

机构信息

National Center of Human Stem Cell Research and Engineering, Institute of Human Reproduction and Stem Cell Engineering, Central South University, Changsha 410078, People's Republic China.

出版信息

Mol Biol Rep. 2009 Apr;36(4):767-74. doi: 10.1007/s11033-008-9243-2. Epub 2008 Apr 4.

Abstract

Methyltransferases play essential roles in modulating important cellular and metabolic processes. A mouse putative N6-DNA methyltransferase gene (GenBank No AY456393) is a novel gene named mN6amt1(mN6A1). To investigate its function in cell fate and protein translation, RNA interference (RNAi)-mediated knock-down method was established. Cell cycle analysis suggests that the cell proliferation decreases after RNAi with mN6A1. The expression plasmid of luciferase was used to detect protein translation, and the results showed that luciferase expression decreased after RNAi with mN6A1, whereas increased after over-expression of mN6A1 or/and eRF1. The binding between mN6A1 and eRF1 was identified by co-immunoprecipitation and pull-down experiments. It might be suggested that mN6A1 participates in protein translation through interaction with eRF1.

摘要

甲基转移酶在调节重要的细胞和代谢过程中发挥着关键作用。小鼠一个假定的N6-DNA甲基转移酶基因(基因库编号AY456393)是一个名为mN6amt1(mN6A1)的新基因。为了研究其在细胞命运和蛋白质翻译中的功能,建立了RNA干扰(RNAi)介导的敲低方法。细胞周期分析表明,用mN6A1进行RNAi后细胞增殖减少。使用荧光素酶表达质粒检测蛋白质翻译,结果显示,用mN6A1进行RNAi后荧光素酶表达降低,而在mN6A1或/和eRF1过表达后增加。通过免疫共沉淀和下拉实验鉴定了mN6A1与eRF1之间的结合。可能提示mN6A1通过与eRF1相互作用参与蛋白质翻译。

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