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We have previously demonstrated (A. H. Batchelor and P. O'Hare, J. Virol. 64:3269-3279, 1990) the selective activity in human neuroblastoma cells (IMR-32) of a promoter located upstream of the latency-associated transcript of herpes simplex virus type 1. In this work, we provide evidence for the basis of the selective activity of this latency-associated promoter (LAP). Recombinant constructs containing sequences up to -143 (relative to the LAP cap site) linked to the chloramphenicol acetyltransferase gene retain strong activity in HeLa cells but exhibit extremely weak activity in IMR-32 cells. Sequences mapping within the 108 bp upstream of -143 to position -251 enhance LAP activity by over 15-fold, restoring optimal levels of expression in IMR-32 cells, but have little or no effect (1.5-fold) in HeLa cells. This cell-type-specific enhancement of promoter activity took place in two major steps, with sequences between -143 and -158 conferring a four- to fivefold effect and sequences between -177 and -251 conferring a further threefold effect. Furthermore, sequences mapping from -40 to -258 could transfer the ability to be expressed in neuroblastoma cells to the normally inactive immediate-early 110K promoter (IE110K), increasing levels of expression by 35-fold. By comparison, this region had a relatively minor effect (twofold) on the activity of the IE110K promoter in HeLa cells, even though this promoter is open to activation by other mechanisms. However, neither of the overlapping subregions from -40 to -143 or -138 to -258 could confer efficient IMR-32 cell expression on the IE110K promoter, and we present alternative models for multiple element requirements or the requirement for a critical site around -140 which is not retained in either subfragment. We provide consistent evidence for a site around -140 and demonstrate the presence selectively in IMR-32 cells of a DNA-binding factor which binds a probe spanning this region. We propose that this element and the cognate factor (IC-1) may be involved in the selective activity of the LAP in neuroblastoma cells.

journal of virology

Localization of cis-acting sequence requirements in the promoter of the latency-associated transcript of herpes simplex virus type 1 required for cell-type-specific activity