Upstream stimulatory factors binding to an E box motif in the R region of the bovine leukemia virus long terminal repeat stimulates viral gene expression

被引:31
作者
Calomme, C
Nguyên, TLA
de Launoit, Y
Kiermer, V
Droogmans, L
Burny, A
Van Lint, C
机构
[1] Free Univ Brussels, Inst Biol & Med Mol, Serv Chim Biol, Lab Virol Mol, B-6041 Gosselies, Belgium
[2] Free Univ Brussels, Fac Med, Microbiol Lab, B-1070 Brussels, Belgium
[3] Inst Pasteur, Inst Biol Lille, CNRS, UMR 8526, F-59021 Lille, France
[4] Free Univ Brussels, Microbiol Lab, Inst Rech CERIA, B-1070 Brussels, Belgium
关键词
D O I
10.1074/jbc.M107441200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The bovine leukemia virus (BLV) promoter is located in its 5'-long terminal repeat and is composed of the U3, R, and U5 regions. BLV transcription is regulated by cis-acting elements located in the U3 region, including three 21-bp enhancers required for transactivation of the BLV promoter by the virus-encoded transactivator Tax(BLV). In addition to the U3 cis-acting elements, both the R and U5 regions contain stimulatory sequences. To date, no transcription factor-binding site has been identified in the R region. Here sequence analysis of this region revealed the presence of a potential E box motif (5'-CACGTG-3'). By competition and supershift gel shift assays, we demonstrated that the basic helix-loop-helix transcription factors USF1 and USF2 specifically interacted with this R region E box motif. Mutations abolishing upstream stimulatory factor (USF) binding caused a reproducible decrease in basal or Tax-activated BLV promoter-driven gene expression in transient transfection assays of B-lymphoid cell lines. Cotransfection experiments showed that the USF1 and USF2a transactivators were able to act through the BLV R region E box. Taken together, these results physically and functionally characterize a USF-binding site in the R region of BLV. This E box motif located downstream of the transcription start site constitutes a new positive regulatory element involved in the transcriptional activity of the BLV promoter and could play an important role in virus replication.
引用
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页码:8775 / 8789
页数:15
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