DeMarco Ricardo, Kowaltowski Andre T, Machado Abimael A, Soares M Bento, Gargioni Cybele, Kawano Toshie, Rodrigues Vanderlei, Madeira Alda M B N, Wilson R Alan, Menck Carlos F M, Setubal João C, Dias-Neto Emmanuel, Leite Luciana C C, Verjovski-Almeida Sergio
Laboratorio de Bioinformatica. Departamento de Bioquimica, Instituto de Quimica, Universidade de São Paulo, 05508-900 São Paulo, Brazil.
J Virol. 2004 Mar;78(6):2967-78. doi: 10.1128/jvi.78.6.2967-2978.2004.
Using the data set of 180,000 expressed sequence tags (ESTs) of the blood fluke Schistosoma mansoni generated recently by our group, we identified three novel long-terminal-repeat (LTR)- and one novel non-LTR-expressed retrotransposon, named Saci-1, -2, and -3 and Perere, respectively. Full-length sequences were reconstructed from ESTs and have deduced open reading frames (ORFs) with several uncorrupted features, characterizing them as possible active retrotransposons of different known transposon families. Alignment of reconstructed sequences to available preliminary genome sequence data confirmed the overall structure of the transposons. The frequency of sequenced transposon transcripts in cercariae was 14% of all transcripts from that stage, twofold higher than that in schistosomula and three- to fourfold higher than that in adults, eggs, miracidia, and germ balls. We show by Southern blot analysis, by EST annotation and tallying, and by counting transposon tags from a Serial Analysis of Gene Expression library, that the four novel retrotransposons exhibit a 10- to 30-fold lower copy number in the genome and a 4- to 200-fold-higher transcriptional rate per copy than the four previously described S. mansoni retrotransposons [corrected]. Such differences lead us to hypothesize that there are two different populations of retrotransposons in S. mansoni genome, occupying different niches in its ecology. Examples of retrotransposon fragment inserts were found into the 5' and 3' untranslated regions of four different S. mansoni target gene transcripts. The data presented here suggest a role for these elements in the dynamics of this complex human parasite genome.
利用我们小组最近生成的180,000个曼氏血吸虫表达序列标签(EST)数据集,我们鉴定出三个新型长末端重复序列(LTR)和一个新型非LTR表达的逆转座子,分别命名为Saci-1、-2和-3以及Perere。从EST重建了全长序列,并推导了具有几个未受损特征的开放阅读框(ORF),将它们表征为不同已知转座子家族的可能活跃逆转座子。将重建序列与可用的初步基因组序列数据进行比对,证实了转座子的整体结构。尾蚴中转座子转录本的测序频率为该阶段所有转录本的14%,比童虫阶段高两倍,比成虫、虫卵、毛蚴和生殖球阶段高三到四倍。我们通过Southern印迹分析、EST注释和计数以及通过对基因表达文库的序列分析计数转座子标签,表明这四个新型逆转座子在基因组中的拷贝数比之前描述的四个曼氏血吸虫逆转座子低10至30倍,每个拷贝的转录率高4至200倍[已修正]。这些差异使我们推测曼氏血吸虫基因组中存在两种不同的逆转座子群体,在其生态中占据不同的生态位。在四个不同的曼氏血吸虫靶基因转录本的5'和3'非翻译区发现了逆转座子片段插入的例子。这里呈现的数据表明这些元件在这种复杂的人体寄生虫基因组动态中发挥作用。