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

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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.

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Acknowledgement

This work was supported by the Hi-Tech Research And Development of China (863 program No. 2006AA02A102), the National Basic Research Program of China (973 program No. 00CB 51010), the National Natural Science Foundation Major Program of China (No. 30570937), the Research Fund for the Doctoral Program of Higher Education from the Ministry of Education of China (No. 20030533002)

Here we especially thank Dr. Adam P. Geballe, Divisions of Human Biology and Clinical Research, Fred Hutchinson Cancer Research Center for kindly giving us the plasmid pEQ842 (containing ORF of eRF1) and Dr. Bogdan Polevoda, Department of biochemical and biophysics, University of Rochester Medical Center for helpful discussion.

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  1. Yongbo Liu

    Present address: Department of Cell and Medical Genetics, Zhengzhou University, Zhengzhou, 450052, People’s Republic China

  2. Dongsong Nie

    Present address: Chemistry and Chemical Engineering Department, Hunan institute of Science and Technology, Yueyang, Hunan, 414000, People’s Republic China

Authors and Affiliations

  1. 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

    Yongbo Liu, Dongsong Nie, Yuelong Huang & Guangxiu Lu

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  1. Yongbo Liu
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  2. Dongsong Nie
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  3. Yuelong Huang
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  4. Guangxiu Lu
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Correspondence to Guangxiu Lu.

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Liu, Y., Nie, D., Huang, Y. et al. RNAi-mediated knock-down of gene mN6A1 reduces cell proliferation and decreases protein translation. Mol Biol Rep 36, 767–774 (2009). https://doi.org/10.1007/s11033-008-9243-2

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  • DOI: https://doi.org/10.1007/s11033-008-9243-2

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