THE JC VIRUS MINIMAL CORE PROMOTER IS GLIAL-CELL SPECIFIC IN-VIVO

The glial cell specificity of the human papovavirus JC (JCV), an etiologic agent for progressive multifocal leukoencephalopathy, is thought to be due to the presence of both positive and negative regulatory elements upstream of the TATA region within the JCV promoter. Here we report that the JCV minimal core promoter, containing only the TATA box and an 8-bp poly(T) region immediately upstream, is sufficient to initiate transcription of an attached gene in glial cells and functions as an autonomously active initiator. We further define the sequences required for this core promoter's glial cell specificity by appropriate substitution and point mutation analysis. Ectopic expression of Tst-1, a POU domain transcription factor that has been implicated in the regulation of oligodendrocyte development, leads to higher activation of the JCV minimal core promoter in Tst-1-deficient glial cells than in non-glial HeLa cells. These results suggest a requirement for a glial cell coactivator(s) for the optimum activation of the JCV minimal core promoter by Tst-1. A discrete affinity of Tst-1 for the JCV core promoter (K-d, 1.4 x 10(-8) M) is also shown to be optimal for its promoter strength. Mutations within the core promoter that maintain this affinity for Tst-1 show maintenance of promoter strength, whereas mutants carrying a change that results in an increased affinity for Tst-1 show reduced transcriptional activity. These results suggest that moderate affinity of Tst-1 for the JCV TATA region may allow the interaction of some glial cell-specific coactivator(s) along with the basal transcription machinery to direct glial cell-specific transcription from the JCV core promoter.