Copeland Claudia S, Heyers Oliver, Kalinna Bernd H, Bachmair Andreas, Stadler Peter F, Hofacker Ivo L, Brindley Paul J
Department of Molecular Parasitology, Institute for Biology, Humboldt University Berlin, Berlin, Germany.
Gene. 2004 Mar 31;329:103-14. doi: 10.1016/j.gene.2003.12.023.
Boudicca is a gypsy-like, long terminal repeat (LTR) retrotransposon that has colonized the genome of the human blood fluke, Schistosoma mansoni. Previous studies have indicated that more than 1000 copies of Boudicca reside within the S. mansoni genome, although many of them may be degenerate and inactive. Messenger RNAs transcribed from genomic copies of Boudicca were investigated by reverse transcription PCR. Overlapping RT-PCR products corresponding to the gag and pol polyproteins of Boudicca, along with relevant sequences of genomic fragments of Boudicca, were assembled into contigs. Consensus sequences from these contigs were used to predict the sequence and structure of transpositionally active copies of the Boudicca retrotransposon. They verified that Boudicca has a kabuki-like Cys-His box motif at the active site of its gag protein, a classic DTG motif as the active site of the protease domain of the pol ORF2, and indicated a contiguous integrase domain at the C-terminus of pol with strong identity to integrase from the LTR retrotransposons CsRn1 and kabuki, as well as to the conserved integrase core domain, GenBank rve (). Models of the secondary structure of the Boudicca transcript suggested that the first AUG was occluded by a stem loop structure, which in turn suggested a method of regulation of expression, at the level of translation, of Boudicca proteins. In addition, phylogenetic analysis targeting discrete domains of Boudicca revealed a generalized radiation in sequences among the multiple copies of Boudicca resident in the schistosome genome.
布迪卡(Boudicca)是一种类似吉普赛人的长末端重复序列(LTR)逆转座子,它已在人类血吸虫曼氏血吸虫(Schistosoma mansoni)的基因组中定殖。先前的研究表明,曼氏血吸虫基因组中存在1000多个布迪卡拷贝,尽管其中许多可能已退化且无活性。通过逆转录PCR研究了从布迪卡基因组拷贝转录的信使RNA。与布迪卡的gag和pol多蛋白相对应的重叠RT-PCR产物,以及布迪卡基因组片段的相关序列,被组装成重叠群。这些重叠群的一致序列被用于预测布迪卡逆转座子转座活性拷贝的序列和结构。他们证实,布迪卡在其gag蛋白的活性位点具有类似歌舞伎的半胱氨酸-组氨酸盒基序,作为pol ORF2蛋白酶结构域活性位点的经典DTG基序,并表明在pol的C末端有一个连续的整合酶结构域,与LTR逆转座子CsRn1和歌舞伎的整合酶具有高度同源性,也与保守的整合酶核心结构域GenBank rve()具有高度同源性。布迪卡转录本二级结构模型表明,第一个AUG被一个茎环结构封闭,这反过来又提示了一种在翻译水平上调控布迪卡蛋白表达的方法。此外,针对布迪卡离散结构域的系统发育分析揭示了血吸虫基因组中多个布迪卡拷贝之间序列的普遍辐射。