The long inverted repeat and the adjacent sequences are major early transcription sites of the human cytomegalovirus genome (M. W. Wathen and M. F. Stinski, J. Virol. 41:462-477, 1982). An early transcription unit which flanks the large terminal repeat was analyzed by RNA mapping at various times after infection. Three unspliced, overlapping RNAs were transcribed from different initiation sites and terminated at the same 3' end. Individual promoters were isolated for all three transcripts. These promoters were activated in trans by viral immediate-early (IE) regulatory proteins after either infection with virus or cotransfection with IE2 alone or IEl plus IE2 genes. DNA sequence analysis detected TATA and CAAT boxes plus multiple-dyad symmetries in the promoter-regulatory region. Deletion analyses showed that the maximum inducible promoter activity lay in a 230-base-pair region. When in the viral genome, the three promoters were regulated differentially during the course of infection. The upstream promoter was used only at late times after infection. Possible reasons for viral RNAs with multiple 5' ends at different times after infection and the recognition of the upstream promoter at only late times after infection are discussed.

A human cytomegalovirus early gene which possesses three temporally regulated promoters is located in the large unique component of the viral genome between 0.054 and 0.064 map units (C.