HIV-1 promotor insertion revealed by selective detection of chimeric provirus-host gene transcripts.
To study host gene activation by retroviral promotor insertion, a polymerase chain reaction (PCR) assay was developed. This method allows a sensitive and selective detection of chimeric provirus-host gene transcripts, hallmarks of insertional activation events, which does not rely on an induction of tumor cell growth. We analysed HIV-1 infected cells of a CD4+ T-cell line (H9), infected peripheral blood mononuclear cells and cells in broncho-alveolar washes of AIDS patients. In each case, a variety of chimeric mRNA molecules were detected using a PCR amplification reaction and 5' primers specific to the HIV-1 LTR and 3' primers specific to poly A of mRNA. In infected H9 lymphocytes, a mRNA was identified encoding a putative protein of 145 amino-acids that was not expressed in uninfected H9 cells. This shows for the first time that HIV-1 can activate transcription of host cellular genes by promotor insertion in a fashion similar to slow-transforming avian and murine retroviruses.
Raineri
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Institute for Medical Microbiology, University of Basel, Switzerland.
Senn
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HP
D000163:Acquired Immunodeficiency Syndrome / Q000382:microbiology; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004279:DNA, Viral; D015967:Gene Expression Regulation, Viral; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic; D011533:Proviruses / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D016662:Virus Integration
D004279:DNA, Viral; D012333:RNA, Messenger