Multiple positive and negative cis-acting elements that mediate transactivation by bel1 in the long terminal repeat...

The bel1 protein of human foamy virus (HFV), a retrovirus, regulates expression of the gene linked to the HFV long terminal repeat (LTR) and is essential for viral gene expression. The mechanism of action of the bel1 protein is unknown, but its action is mediated through the U3 region of the LTR. To determine which U3 sequences are critical for transactivation by bel1, a series of hybrid vectors consisting of a mutant HFV LTR and the chloramphenicol acetyltransferase gene were constructed and tested for their responsiveness to the bel1 protein by using transient assays after transfection. The target sequences for transactivation by bel1 were mapped to five regions in the U3 domain of the LTR: nucleotides -559 to -506, -454 to -418, -360 to -342, -327 to -284, and -116 to -89 (+1 represents the transcription initiation site). No significant sequence similarity was identified among the five target sites. The observation that the multiple distinct elements in the HFV LTR are the targets for bel1 transactivation is different from observations with other human retroviral systems. The regulation mechanism of HFV bel1 protein-mediated transactivation appears to be analogous to that of some DNA virus transactivators that increase transcription from numerous different viral promoters with little sequence similarity shared among them. We demonstrated that multiple bel1-responsive elements (BRE) can act as bel1-dependent enhancer elements, while a single copy of one BRE, BREe, can serve as an upstream activating element in both orientations. In addition, the region between -466 and -498 was identified as responsible for the downregulation of gene expression directed by BREa, which requires its upstream sequence element to act as a bel1-dependent enhancer element in a heterologous promoter.