A GENERAL-METHOD FOR THE IDENTIFICATION OF TRANSCRIBED RETROVIRUS SEQUENCES (R-U5 PCR) REVEALS THE EXPRESSION OF THE HUMAN ENDOGENOUS RETROVIRUS LOCI HERV-H AND HERV-K IN TERATOCARCINOMA CELLS
A GENERAL-METHOD FOR THE IDENTIFICATION OF TRANSCRIBED RETROVIRUS SEQUENCES (R-U5 PCR) REVEALS THE EXPRESSION OF THE HUMAN ENDOGENOUS RETROVIRUS LOCI HERV-H AND HERV-K IN TERATOCARCINOMA CELLS
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DOI：
10.1006/viro.1993.1066
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发表时间：
1993-02-01
期刊：
影响因子：
3.7
通讯作者：
KURTH, R
中科院分区：
文献类型：
--
作者：
LOWER, R;LOWER, J;KURTH, R
在过去的十年中，不同类型的内源性逆转录病毒序列已被确定在人类基因组中，通常通过低严格性杂交采用进化保守的动物逆转录病毒基因的DNA探针。尽管发现测序的所有人类基因组基因座都有缺陷或散布有终止密码子，但间接证据正在积累，即人类内源性逆转录病毒基因座至少在某些情况下表达。一个例子是通过电子显微镜观察到的人畸胎瘤细胞系中逆转录病毒颗粒的合成。为了建立病毒表达和基因组位点之间的联系，我们使用普遍适用的方法在人细胞mRNA群体中搜索逆转录病毒RNA。通过逆转录延伸与推定的逆转录病毒引物结合位点互补的tRNA衍生的引物，并且用均聚物延伸并通过PCR（R-U 5 PCR）扩增该产物。这些产物的克隆和测序显示，内源性逆转录病毒基因座HERV-H和HERV-K在产生逆转录病毒颗粒的人畸胎瘤细胞系中表达。在北方印迹中检测到的四种HERV-K mRNA的大小分布使人联想到在许多外源性逆转录病毒中观察到的复杂表达模式。
During the past decade, different types of endogenous retroviral sequences have been defined in the human genome usually by low stringency hybridization employing DNA probes of evolutionary conserved animal retrovirus genes. Although all human genomic loci sequenced were found to be defective or interspersed with stop codons, indirect evidence is accumulating that human endogenous retroviral loci are expressed at least in some instances. One example is the synthesis of retroviral particles in human teratocarcinoma cell lines observed by electron microscopy. To establish a link between virus expression and genomic loci we searched for retroviral RNA in human cellular mRNA populations using a generally applicable method. A tRNA-derived primer complementary to a putative retroviral primer binding site was extended by reverse transcription and this product was elongated with a homopolymeric stretch and amplified by PCR (R-U5 PCR). Cloning and sequencing of such products revealed that the endogenous retroviral loci HERV-H and HERV-K are expressed in those human teratocarcinoma cell lines which produce retroviral particles. The size distribution of four HERV-K mRNAs detected in Northern blots is reminiscent of the complex expression pattern seen with a number of exogenous retroviruses.