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Molecular characterisation of RecQ homologues in Arabidopsis thaliana

机译:拟南芥中RecQ同源物的分子表征

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摘要

Members of the RecQ family of DNA helicases are involved in processes linked to DNA replication, DNA recombination and gene silencing. RecQ homologues of various animals have been described recently. Here, for the first time for plants, we characterised cDNAs of all in all six different RecQ-like proteins that are expressed to different extents in Arabidopsis thaliana. Surprisingly, three of these proteins are small in size [AtRecQl1, AtRecQl2, AtRecQl3—606, 705 and 713 amino acids (aa), respectively], whereas the two bigger proteins result from a duplication event during plant evolution [AtRecQl4A and AtRecQl4B—1150 and 1182 aa, respectively]. Another homologue (AtRecQsim, 858 aa) most probably arose by insertion of an unrelated sequence within its helicase domain. The presence of these homologues demonstrates the conservation of RecQ family functions in higher eukaryotes. We also detected a small gene (AtWRNexo) encoding 285 aa which, being devoid of any RecQ-like helicase domain, reveals a striking homology to the exonuclease domain of human Werner protein, a prominent RecQ helicase of larger size. By means of the two-hybrid assay we were able to detect an interaction between AtWRNexo and AtRecQl2, indicating that activities that reside in a single protein chain in mammals might in plants be complemented in trans.
机译:RecQ系列DNA解旋酶的成员参与与DNA复制,DNA重组和基因沉默相关的过程。最近已经描述了各种动物的RecQ同源物。在这里,对于植物,我们首次表征了在拟南芥中以不同程度表达的所有六个不同RecQ样蛋白的全部cDNA。出乎意料的是,这些蛋白质中的三种蛋白质的大小很小[分别为AtRecQl1,AtRecQl2,AtRecQl3-606、705和713个氨基酸(aa)],而两种较大的蛋白质是植物进化过程中发生的重复事件产生的[AtRecQl4A和AtRecQl4B-1150和1182 aa]。另一个同源物(AtRecQsim,858aa)很可能是通过在其解旋酶结构域中插入无关序列而引起的。这些同源物的存在证明了在高等真核生物中RecQ家族功能的保守性。我们还检测到一个编码285aa的小基因(AtWRNexo),该基因不含任何类似RecQ的解旋酶结构域,揭示了与人Werner蛋白外切核酸酶结构域的惊人同源性,这是一种较大尺寸的著名RecQ解旋酶。通过双杂交测定,我们能够检测到AtWRNexo与AtRecQ12之间的相互作用,这表明存在于哺乳动物单条蛋白链中的活性可能在植物中反式互补。

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