Departamento de Biología Funcional, Área de Genética. Universidad de Oviedo, 33071 Oviedo, Spain.
Departamento de Biología Funcional, Área de Genética. Universidad de Oviedo, 33071 Oviedo, Spain.
Gene. 2020 May 30;741:144546. doi: 10.1016/j.gene.2020.144546. Epub 2020 Mar 9.
Retrotransposon roo is one of the most active elements in Drosophila melanogaster. The level of nucleotide diversity between copies of roo is very low but structural variation in the 5'-UTR is considerable. Transposition of roo at high frequency (around 5 × 10 per generation) has been shown previously in the set of mutation accumulation lines named Oviedo. Here we isolated thirteen individual insertions by inverse PCR and sequenced the 5' end of the elements (between 1663 and 2039 nt) including the LTR, the 5'-UTR and a fragment of 661 nucleotides from the ORF, to study whether the new transposed copies come from a unique variant (the master copy model) or different elements are able to move (the transposon model). The elements in the Oviedo lines presented the same structural variants as the reference genome. Different structural variants were active, a behaviour compatible with the "transposon model" in which the copies localized in multiple sites in the genome are able to transpose. At the level of sequence, the copies of roo in our lines are highly similar to the elements in the reference genome. The phylogenetic tree shows a shallow diversification with unsupported nodes denoting that all the elements currently active are very young. This observation together with the great polymorphism in insertion sites implies a rapid turnover of the elements.
反转录转座子 roo 是黑腹果蝇中最活跃的元件之一。roo 拷贝之间的核苷酸多样性水平非常低,但 5'-UTR 中的结构变异相当大。先前在名为 Oviedo 的突变积累系中已经证明了 roo 的高频转座(约每代 5×10)。在这里,我们通过反向 PCR 分离了 13 个个体插入,并对元件的 5'端进行了测序(1663 到 2039 个核苷酸),包括 LTR、5'-UTR 和 ORF 的 661 个核苷酸片段,以研究新转座的拷贝是否来自独特的变体(主拷贝模型)或不同的元件能够移动(转座子模型)。Oviedo 系中的元件呈现出与参考基因组相同的结构变体。不同的结构变体是活跃的,这与“转座子模型”一致,其中定位在基因组多个位置的拷贝能够转座。在序列水平上,我们系中的 roo 拷贝与参考基因组中的元件高度相似。系统发育树显示出浅层多样化,没有支持的节点表示当前活跃的所有元件都非常年轻。这一观察结果以及插入位点的高度多态性表明元件的快速更替。