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非自主I型内含子逆转座子的翻译是转座活性的关键决定因素:对逆转座子介导的基因组进化的启示。

Translation from nonautonomous type IAP retrotransposon is a critical determinant of transposition activity: implication for retrotransposon-mediated genome evolution.

作者信息

Saito Ei-Suke, Keng Vincent W, Takeda Junji, Horie Kyoji

机构信息

Department of Social and Environmental Medicine, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.

出版信息

Genome Res. 2008 Jun;18(6):859-68. doi: 10.1101/gr.069310.107. Epub 2008 May 2.

Abstract

Retrotransposons constitute a major component of the genome and their proliferation significantly impacts genome evolution. Retrotransposons can propagate autonomously or nonautonomously. Nonautonomous type transposition occurs through trans-complementation by autonomous type retrotransposons. While autonomous type retrotransposons have been studied extensively, the translation products from nonautonomous type retrotransposons are not well characterized. In a previous study, we isolated both autonomous and nonautonomous type intracisternal A particle (IAP) elements from the mouse genome and established a tissue culture assay to examine trans-complementation of nonautonomous type IAP element. Using this system in the present study, we determined an active role for the translation product from nonautonomous type IAP element. Point mutations that either eliminated or truncated the IAP protein were introduced and their effects on trans-complementation were examined. Trans-complementation efficiency correlated with the expression of nonautonomous type IAP protein. The effect of nonautonomous type IAP protein was observed only when it was provided in cis, suggesting an interaction of nonautonomous type IAP protein and its transcript immediately after transcription. Interaction of autonomous and nonautonomous type IAP proteins was demonstrated by immunostaining and coimmunoprecipitation assay. Based on these findings, we propose a model in which nonautonomous type IAP protein associates with its transcript, recruits autonomous type IAP protein, and promotes the assembly of transposition competent IAP particle. The active role of the nonautonomous type IAP protein revealed in this study may provide a new insight into retrotransposon proliferation within the genome.

摘要

逆转录转座子是基因组的主要组成部分,它们的增殖对基因组进化有重大影响。逆转录转座子可以自主或非自主地传播。非自主型转座通过自主型逆转录转座子的反式互补作用发生。虽然自主型逆转录转座子已得到广泛研究,但非自主型逆转录转座子的翻译产物尚未得到很好的表征。在先前的一项研究中,我们从小鼠基因组中分离出自主型和非自主型的胞内A颗粒(IAP)元件,并建立了一种组织培养检测方法来检测非自主型IAP元件的反式互补作用。在本研究中使用该系统,我们确定了非自主型IAP元件翻译产物的积极作用。引入了消除或截断IAP蛋白的点突变,并检测了它们对反式互补作用的影响。反式互补效率与非自主型IAP蛋白的表达相关。仅当非自主型IAP蛋白以顺式提供时才观察到其作用,这表明非自主型IAP蛋白与其转录本在转录后立即发生相互作用。通过免疫染色和免疫共沉淀分析证明了自主型和非自主型IAP蛋白之间的相互作用。基于这些发现,我们提出了一个模型,其中非自主型IAP蛋白与其转录本结合,招募自主型IAP蛋白,并促进具有转座能力的IAP颗粒的组装。本研究中揭示的非自主型IAP蛋白的积极作用可能为基因组内逆转录转座子的增殖提供新的见解。

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