The enhancer of JC virus restricts its gene expression to the brain. In a previous study, we demonstrated an enhancer element, nuclear factor I (NFI) motif, in the middle of the enhancer. Here, we demonstrate that the NFI motif is a tissue-specific element. We further present a new tissue-specific element, SacI motif, just upstream from the NFI motif. These motifs showed transcriptional enhancement both and and acted upon a heterologous adenovirus major late promoter. DNase I footprint analyses demonstrated that the SacI motif bound to a brain nuclear factor, and that its binding region overlapped with the NFI motif. Gel shift experiments with the SacI motif revealed that the populations of SacI motif factors in the brain and HeLa cell extracts were different. Together with our previous findings about tissue-specific NFI-like factor(s), cooperation of NFI motif and SacI motif factors may be required for the strong brain specificity of the viral gene expression.


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