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.
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.
Löwer
R
R
Paul-Ehrlich-Institut, Langen/Frankfurt, Germany.
Löwer
J
J
Tondera-Koch
C
C
Kurth
R
R
D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D002822:Choriocarcinoma / Q000235:genetics / Q000382:microbiology; D015342:DNA Probes; D015751:Genes, pol; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D012339:RNA, Ribosomal, 28S / Q000235:genetics; D012343:RNA, Transfer / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D012190:Retroviridae / Q000235:genetics* / Q000302:isolation & purification*; D012689:Sequence Homology, Nucleic Acid; D013724:Teratoma / Q000235:genetics / Q000382:microbiology*; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D015342:DNA Probes; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D012339:RNA, Ribosomal, 28S; D012367:RNA, Viral; D012343:RNA, Transfer