Expression of intracisternal A-particle-related retroviral element-encoded envelope proteins detected in cell lines.
Intracisternal A-particle (IAP) retrotransposons of rodents express gag and pol proteins for assembly of intracellular viruslike particles but lack an env gene. The recently described IAP-related family of retroviral elements contains a reading frame with close resemblance to retroviral env genes (IAPEs) (F. U. Reuss and H. C. Schaller, J. Virol. 65:5702-5709, 1991). I now report the analysis of cellular IAPE mRNAs and detection of IAPE env proteins. IAPE elements are transcribed in cell lines NH15-CA2 and AtT20. Four major transcripts of 4.2, 3.9, 2.8, and 1.3 kb are detected and characterized by probes specific for defined regions of the cloned IAPE-1 cDNA. The 2.8-kb mRNA is shown to lack gag and pol genes but comprises an env gene and U3 region, as expected for a subgenomic env mRNA. Polymerase chain reaction amplification and cloning of such mRNAs confirmed the absence of gag and pol genes 5' from the env gene and implicates env mRNA generation by a splicing event. A polyclonal anti-IAPE env antiserum, raised against a bacterial IAPE-env fusion protein, specifically detects N-glycosylated env proteins of 91 kDa or less in cell lines positive for IAPE mRNA. IAPE env proteins of different sizes represent independent translation products. After inhibition of N-glycosylation, env proteins in the size predicted from the env gene sequence or smaller are present. These results provide evidence that putative IAPE env proteins are synthesized in vivo. Envelope protein expression by an IAP-related retroviral element identifies IAPEs as a possible missing link between IAP retrotransposons and retroviruses.
Reuss
F U
FU
Zentrum für Molekulare Biologie, Universität Heidelberg, Germany.
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004279:DNA, Viral; D015686:Gene Products, env / Q000235:genetics*; D007426:Genes, Intracisternal A-Particle / Q000235:genetics*; D015750:Genes, gag; D015751:Genes, pol; D006031:Glycosylation; D051379:Mice; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012326:RNA Splicing; D012360:RNA, Transfer, Phe / Q000378:metabolism; D015531:Radioimmunoprecipitation Assay; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D004279:DNA, Viral; D015686:Gene Products, env; D012360:RNA, Transfer, Phe