TRANSLATION OF LINE-1 DNA ELEMENTS INVITRO AND IN HUMAN-CELLS
TRANSLATION OF LINE-1 DNA ELEMENTS INVITRO AND IN HUMAN-CELLS
复制标题
DOI：
10.1073/pnas.87.18.6990
复制
发表时间：
1990-09-01
影响因子：
11.1
通讯作者：
FANNING, TG
中科院分区：
文献类型：
--
作者：
LEIBOLD, DM;SWERGOLD, GD;FANNING, TG
在人类基因组中发现的LINE-1(L1)散布DNA序列家族(L1 Homosapiens，L1Hs)包括活性转座元件。目前转座机制的模型包括RNA中间体的逆转录和利用元件编码的蛋白质。我们报道，针对L1Hs 5‘’开放阅读框架(ORF1)编码的多肽的抗血清在人类细胞中检测到内源性ORF1蛋白以及适当的转基因重组载体的ORF1产物。在所测试的细胞系中，内源性多肽在畸胎癌和绒毛膜癌细胞中含量最高；它似乎是.apprxeq的单一物种。38 kDa。相比之下，由代表全长、多腺化细胞质L1Hs RNA的cDNA在体外合成的RNA，在体外翻译时，ORF1产物的大小略有不同。这与不同的cDNA是不同的，代表不同基因组L1Hs元件的转录是一致的。体外研究还表明，ORF1的翻译是在8月的第一个密码子开始的。最后，无缘无故地检测到ORF1-ORF2融合蛋白。
The LINE-1 (L1) family of interspersed DNA sequences found throughout the human genome (L1 Homosapiens, L1Hs) includes active transposable elements. Current models for the mechanism of transposition involve reverse transcription of an RNA intermediate and utilization of element-encoded proteins. We report that an antiserum against the polypeptide encoded by the L1Hs 5'' open reading frame (ORF1) detects, in human cells, an endogenous ORF1 protein as well as the ORF1 product of an appropriate transfecting recombinant vector. The endogenous polypeptide is most abundant in teratocarcinoma and choriocarcinoma cells, among those cell lines tested; it appears to be a single species of .apprxeq. 38 kDa. In contrast, RNAs synthesized in vitro from cDNAs representing full-length, polyadenylated cytoplasmic L1Hs RNA yield, upon in vitro translation, ORF1 products of slightly different sizes. This is consistent with the fact that the various cDNAs are different and represent transcription of different genomic L1Hs elements. In vitro studies additionally suggest the translation of ORF1 is initiated at the first AUG codon. Finally, in no cause was an ORF1-ORF2 fusion protein detected.