Feldbrügge M, Arizti P, Sullivan M L, Zamore P D, Belasco J G, Green P J
MSU-DOE Plant Research Laboratory, Michigan State University, East Lansing 48824-1312, USA.
Plant Mol Biol. 2002 May;49(2):215-23. doi: 10.1023/a:1014936824187.
The labile SAUR transcripts from higher plants contain a conserved DST sequence in their 3'-untranslated regions. Two copies of a DST sequence from soybean are sufficient to destabilize reporter transcripts in cultured tobacco cells whereas variants bearing mutations in the conserved ATAGAT or GTA regions are inactive. To investigate the potential for conserved recognition components in mammalian and plant cells, we examined the function of this instability determinant in mouse NIH3T3 fibroblasts and tobacco BY2 cells. In fibroblasts, a tetrameric DST element from soybean accelerated deadenylation and decay of a reporter transcript. However, a version mutated in the ATAGAT region was equally effective in this regard, and a tetrameric DST element from Arabidopsis was inactive. In contrast, the soybean DST element was more active as an mRNA instability element than the mutant version and the Arabidopsis element, when tested as tetramers in tobacco cells. Hence, the plant DST element is not recognized in animal cells with the same sequence requirements as in plant cells. Therefore, its mode of recognition appears to be plant-specific.
高等植物中不稳定的SAUR转录本在其3'非翻译区含有保守的DST序列。来自大豆的两份DST序列足以使培养的烟草细胞中的报告基因转录本不稳定,而在保守的ATAGAT或GTA区域带有突变的变体则无活性。为了研究哺乳动物和植物细胞中保守识别元件的可能性,我们检测了这种不稳定决定因素在小鼠NIH3T3成纤维细胞和烟草BY2细胞中的功能。在成纤维细胞中,来自大豆的四聚体DST元件加速了报告基因转录本的去腺苷酸化和衰变。然而,在ATAGAT区域发生突变的一个版本在这方面同样有效,而来自拟南芥的四聚体DST元件则无活性。相比之下,当在烟草细胞中作为四聚体进行测试时,大豆DST元件作为mRNA不稳定元件比突变版本和拟南芥元件更具活性。因此,植物DST元件在动物细胞中不能像在植物细胞中那样以相同的序列要求被识别。因此,其识别模式似乎是植物特异性的。