Regulation of the Long Terminal Repeat in Visna Virus by a Transcription Factor Related to the AML/PEBP2/CBF Superfamily

• Published in: Virology (New York, N.Y.) N.Y.), 1997-03, Vol.229 (1), p.240-250
• Main Authors: Sutton, Keith A., Lin, Chung-Tien, Harkiss, Gordon D., Mcconnell, Ian, Sargan, David R.
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Cartilage - cytology; Cartilage - virology; Cells, Cultured; Chloramphenicol O-Acetyltransferase - genetics; DNA, Viral; Genes, Reporter; Molecular Sequence Data; Promoter Regions, Genetic; Protein Binding; Repetitive Sequences, Nucleic Acid; Sequence Homology, Nucleic Acid; Sheep; Transcription Factors - metabolism; VIRUS VISNA MAEDI; Visna-maedi virus - genetics
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1006/viro.1996.8432

The long terminal repeats of maedi visna virus strain 1514 contain a consensus AP-1 binding site which has been shown to be important in controlling virus transcription. However, this consensus site is absent in strain EV-1. Here, we have compared the ability of oligonucleotides corresponding to LTR sequences from EV-1 with those from 1514 to bind transcription factors in competitive gel retardation assays and activate reporter gene expression. The experiments demonstrated no observable binding of AP-1 to the EV-1-derived sequences and significant differences in the abilities of the 1514 and EV-1 sequences to activate transcription. However, both viral sequences interacted with a second, previously undetected, transcription factor. This factor gave specific gel shifts which were competed by an oligonucleotide containing the consensus sequence for the AML/PEBP2/CBF family of transcriptional factors, but not by control AP-1 or OCT-1 oligonucleotides. The factor was therefore denoted AML (vis). A second AML (vis) site, noted upstream of the TATA box proximal AP-1 site, gave single shifts which were competed by the downstream AML (vis) oligonucleotide. Both sites were functional in transfection assays. In gel shift retardation assays, polyclonal antisera directed against known runt domain proteins were able to supershift part of the AML (vis) binding activity in nuclear extracts from physiologically relevant cell types. The results thus suggest that the AML (vis) binding factor belongs to the AML/PEBP2/CBF family of transcription factors and may be important in controlling virus replication in these and other strains of ruminant lentiviruses.