Willhoeft Ute, Buss Heidrun, Tannich Egbert
Bernhard Nocht Institute for Tropical Medicine, 20359 Hamburg, Germany.
Infect Immun. 2002 Dec;70(12):6798-804. doi: 10.1128/IAI.70.12.6798-6804.2002.
While comparing gene expression in the pathogenic organism Entamoeba histolytica and the closely related but nonpathogenic species Entamoeba dispar, we discovered that the E. histolytica abundant polyadenylated transcript 2 (ehapt2) and corresponding genomic copies are absent in E. dispar. Although polyadenylated, ehapt2 does not contain any overt open reading frame. Southern blot and sequence analyses revealed that about 500 copies of ehapt2 genomic elements were present in each cell and that the copies were distributed throughout the ameba genome. The various ehapt2 elements are regularly located in the vicinity of protein-encoding genes, downstream of pyrimidine-rich sequence stretches (40 to 125 bp; CT content, 79.2 to 85.5%), and are flanked by duplicated target sites of variable length. Target site duplications were obviously generated during integration of ehapt2 into the E. histolytica genome as one copy of the flanking repeat and the complete ehapt2 element are specifically absent in orthologous E. dispar genomic sequences. ehapt2 shares 3' sequences with EhRLE, a recently identified non-long-terminal-repeat (non-LTR) retrotransposon-like element of E. histolytica, which contains a conceptual open reading frame for reverse transcriptase. Thus, ehapt2 has all of the properties of nonautonomous non-LTR retrotransposons. A comparison of various E. histolytica isolates suggested that transposition of ehapt2 takes place at a very low frequency as the genomic localization of ehapt2 elements was found to be well conserved. A mobile element such as ehapt2 could be a suitable mechanism to explain the infrequent and late transition of E. histolytica from a harmless gut commensal to an invasive pathogen.
在比较致病生物溶组织内阿米巴和密切相关但无致病性的物种迪氏内阿米巴的基因表达时,我们发现迪氏内阿米巴中不存在溶组织内阿米巴丰富的聚腺苷酸化转录本2(ehapt2)及其相应的基因组拷贝。尽管ehapt2是聚腺苷酸化的,但它不包含任何明显的开放阅读框。Southern印迹和序列分析表明,每个细胞中存在约500个ehapt2基因组元件拷贝,并且这些拷贝分布在整个阿米巴基因组中。各种ehapt2元件通常位于蛋白质编码基因附近,在富含嘧啶的序列片段(40至125 bp;CT含量,79.2至85.5%)下游,并且两侧是长度可变的重复靶位点。靶位点重复显然是在ehapt2整合到溶组织内阿米巴基因组过程中产生的,因为在直系同源的迪氏内阿米巴基因组序列中,侧翼重复序列的一个拷贝和完整的ehapt2元件特异性缺失。ehapt2与EhRLE共享3'序列,EhRLE是最近在溶组织内阿米巴鉴定出的一种非长末端重复(non-LTR)逆转座子样元件,它包含一个逆转录酶的概念性开放阅读框。因此,ehapt2具有非自主非LTR逆转座子的所有特性。对各种溶组织内阿米巴分离株的比较表明,ehapt2的转座频率非常低,因为发现ehapt2元件的基因组定位非常保守。像ehapt2这样的移动元件可能是解释溶组织内阿米巴从无害的肠道共生菌向侵袭性病原体罕见且晚期转变的合适机制。