Wilkinson D A, Freeman J D, Goodchild N L, Kelleher C A, Mager D L
Terry Fox Laboratory, B.C. Cancer Research Centre, Vancouver, Canada.
J Virol. 1990 May;64(5):2157-67. doi: 10.1128/JVI.64.5.2157-2167.1990.
Northern (RNA) blot analysis of RNA from various human cell lines and tissues has demonstrated that elements belonging to the RTVL-H family of human endogenous retroviruslike sequences are expressed in several cell types. The highest levels of RTVL-H-related RNAs were observed in teratocarcinoma cell line NTera2D1, HeLa cells, two bladder carcinoma cell lines, and normal amniotic tissue. Expression was also observed in normal chorion and in some other cell lines. The RTVL-H transcription pattern varied among the different cell types, but several expressed a unit-length 5.6-kilobase transcript. Characterization of cDNA clones corresponding to transcripts present in NTera2D1 cells indicates that the complex transcription pattern observed in these cells is generated by the following: (i) transcription of both full-length and deleted genomic elements, which is initiated within the 5' long terminal repeat (LTR) and, in all but one case, polyadenylated in the 3' LTR; (ii) the splicing of both unit-length transcripts and transcripts from a deleted element; (iii) transcription involving solo LTR sequences; and (iv) transcription which, in one case, reads through the 3' LTR into flanking cellular sequences. Sequence data obtained from 25 cDNA clones revealed that at least 13 RTVL-H elements are expressed in NTera2D1 cells. The positions of several termination codons within the pol region are the same among nine different elements, indicating that an ancestral RTVL-H element bearing these mutations dispersed within the genome. We also found that RTVL-H expression varied among samples of amnion and chorion tissue isolated from different individuals. These findings demonstrate that regulated autonomous expression of RTVL-H sequences occurs in human cells.
对来自各种人类细胞系和组织的RNA进行的Northern(RNA)印迹分析表明,属于人类内源性逆转录病毒样序列RTVL-H家族的元件在几种细胞类型中表达。在畸胎瘤细胞系NTera2D1、HeLa细胞、两种膀胱癌细胞系和正常羊膜组织中观察到最高水平的RTVL-H相关RNA。在正常绒毛膜和其他一些细胞系中也观察到了表达。RTVL-H的转录模式在不同细胞类型中有所不同,但有几种细胞表达了一个单位长度的5.6千碱基转录本。对与NTera2D1细胞中存在的转录本相对应的cDNA克隆进行的表征表明,在这些细胞中观察到的复杂转录模式是由以下因素产生的:(i)全长和缺失的基因组元件的转录,其在5'长末端重复序列(LTR)内起始,除了一个例外,在3'LTR中进行多聚腺苷酸化;(ii)单位长度转录本和缺失元件转录本的剪接;(iii)涉及单独LTR序列的转录;(iv)在一种情况下,转录通读3'LTR进入侧翼细胞序列。从25个cDNA克隆获得的序列数据显示,至少有13个RTVL-H元件在NTera2D1细胞中表达。在九个不同元件中,pol区域内几个终止密码子的位置相同,这表明携带这些突变的祖先RTVL-H元件在基因组中分散。我们还发现,从不同个体分离的羊膜和绒毛膜组织样本中RTVL-H的表达有所不同。这些发现表明,RTVL-H序列在人类细胞中发生受调控的自主表达。