Journal
JOURNAL OF VIROLOGY
Volume 78, Issue 23, Pages 12788-12799Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/JVI.78.23.12788-12799.2004
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Funding
- NIAID NIH HHS [R21 AI013562, R01 AI013562, AI-13562] Funding Source: Medline
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The human cytomegalovirus (CMV) enhancer has a distal component (positions -550 to -300) and a proximal component (-300 to -39) relative to the transcription start site (+ 1) of the major immediate-early (MIE) promoter. Without the distal enhancer, human CMV replicates slower and has a small-plaque phenotype. We determined the sequence requirements of the proximal enhancer by making 5'-end deletions to positions -223, -173, -116, -67, and -39. Even though recombinant virus with the proximal enhancer deleted to -39 has the minimal TATA box-containing MIE promoter element, it cannot replicate independently in human fibroblast cells. Recombinant virus with a deletion to -67 has an Sp-1 transcription factor binding site which may represent a minimal enhancer element for recombinant virus replication in human fibroblast cells. Although recombinant virus with a deletion to -223 replicates to titers at least 100-fold less than that of the wild-type virus, it replicates to titers 8-fold higher than that of recombinant virus with a deletion to - 173 and 20-fold higher than that of virus with a deletion to -67. Recombinant virus with a deletion to -173 replicates more efficiently than that with a deletion to -116. There was a direct correlation between the level of infectious virus replication and time after infection, amount of MIE gene transcription, MIE and early viral protein synthesis, and viral DNA synthesis. The extent of the proximal enhancer determines the efficiency of viral replication.
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