Maestre J, Tchénio T, Dhellin O, Heidmann T
Unité de Physicochimie et Pharmacologie des Macromolécules Biologiques, CNRS URA 147, Institut Gustave Roussy, Villejuif, France.
EMBO J. 1995 Dec 15;14(24):6333-8. doi: 10.1002/j.1460-2075.1995.tb00324.x.
Using a sensitive assay for detection of reverse transcription events, we demonstrate that human HeLa cells can 'retropose', i.e. reverse transcribe and integrate, the mRNA of a naive reporter gene, at a low but detectable frequency. Furthermore, we show that the retroposed copies have all the hallmarks of the processed pseudogenes naturally found in the mammalian genome: they lack intron and 5' promoter sequence, they have acquired a 3' poly(A) tail, and they are flanked by short repeats (< 15 bp) of target DNA sequence. These results demonstrate that human cells possess an endogenous reverse transcription activity, which is not restricted to transcripts of transposable elements, and which is likely to be involved in the formation, still ongoing, of a large fraction of the eukaryotic genome.
通过使用一种灵敏的检测方法来检测逆转录事件,我们证明人类HeLa细胞能够以低但可检测到的频率“逆转座”,即逆转录并整合一个原始报告基因的mRNA。此外,我们表明逆转座的拷贝具有哺乳动物基因组中天然存在的加工假基因的所有特征:它们缺乏内含子和5'启动子序列,获得了3'多聚腺苷酸尾,并且两侧是靶DNA序列的短重复序列(<15 bp)。这些结果表明人类细胞具有内源性逆转录活性,这种活性不仅限于转座元件的转录本,并且可能参与了仍在进行的大部分真核基因组的形成。