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Prior studies have demonstrated that a small proportion of blood lymphocytes from patients with human cytomegalovirus (HCMV) infection express only the viral immediate-early (IE) genes (L. Einhorn and A. Ost, J. Infect. Dis. 149:207-214, 1984; G. P. A. Rice, R. D. Schrier, and M. B. A. Oldstone, Proc. Natl. Acad. Sci. USA 81:6134-6138, 1984). The present studies demonstrate that the IE genes of HCMV are transcribed in Jurkat cells (T lymphocytes) only after activation of the cells with mitogens. Transcription of the IE genes is from an upstream enhancer promoter-regulatory region containing several different repeated sequence motifs. Chimeric plasmids were constructed with just a single copy or three copies of a synthetic oligonucleotide sequence of either the 16-, 18-, 19-, or 21-base-pair (bp) repeat elements upstream of the minimal wild-type promoter sequence to drive expression of the indicator gene, chloramphenicol acetyltransferase (CAT). The 18- or 19-bp motifs in the enhancer region were found to be important in mediating the effect of the mitogens. However, the CAT activity detected with the 19-bp repeat was always significantly higher than that found with the 18-bp repeat. There was an additive effect by multiple copies of the 18- or 19-bp repeat sequences on gene expression. The 19-bp repeat contains a sequence identical to that described for a cyclic AMP (cAMP) response element, and plasmids containing only this sequence and the minimal promoter sequences upstream of the CAT gene respond to agents which increase intracellular cAMP. Functional cAMP response elements are present in the wild-type promoter-regulatory region and are associated with the 19-bp repeat sequences. It is proposed that activation of lymphocytes results in expression of the IE genes of HCMV, in part via the activation of cellular trans-acting factors which interact with the 18- and 19-bp motifs in the HCMV IE promoter-regulatory region. The 19-bp repeat is the major contributor to the strength of this enhancer-containing promoter-regulatory region.

journal of virology

The promoter-regulatory region of the major immediate-early gene of human cytomegalovirus responds to T-lymphocyte stimulation and contains functional cyclic AMP-response elements