Analysis of the role of the bel and bet open reading frames of human foamy virus by using a new quantitative assay.
We have constructed a BHK-21-derived indicator cell line containing a single integrated copy of the beta-galactosidase (beta-Gal) gene under control of the human foamy virus (HFV) long terminal repeat promoter (from -533 to +20). These foamy virus-activated beta-Gal expression (FAB) cells can be used in a quantitative assay to measure the infectious titer of HFV. Our results show that the FAB assay is 50 times more sensitive than determination of the virus titer by the end-point dilution method. Using the FAB assay, we have found that HFV can productively replicate in several erythroblastoid cell lines as well as in the Jurkat T-cell line. We have also examined the roles of bel2, bet, and bel3 in viral replication by constructing proviral HFV clones in which the reading frame of Bel2, Bet, or Bel3 is disrupted by placement of translation stop codons. Analysis of these mutants reveals that while the bel3 gene is not required for viral replication in vitro, mutations in the bel2 or bet gene decrease cell-free viral transmission approximately 10-fold.
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Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104-2092.
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D001483:Base Sequence; D002460:Cell Line; D005814:Genes, Viral; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D012091:Repetitive Sequences, Nucleic Acid; D012191:Retroviridae Proteins / Q000235:genetics*; D016092:Spumavirus / Q000235:genetics*; D015534:Trans-Activators; D015678:Viral Structural Proteins / Q000235:genetics*; D014779:Virus Replication
D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D012191:Retroviridae Proteins; D015534:Trans-Activators; D015678:Viral Structural Proteins