The human cytomegalovirus immediate early enhancer-promoter is responsive to activation by the adenovirus-5 13S E1A gene.
The stimulatory effects of the 13S adenovirus E1A gene product on the human cytomegalovirus (HCMV) major immediate early (IE) enhancer were examined. Chimeric plasmids containing cloned portions of the HCMV major IE enhancer-promoter positioned upstream of the chloramphenicol acetyltransferase gene (cat) were cotransfected into HeLa cells with the plasmid p13S-wt which contained a cDNA encoding the adenovirus 13S E1A gene product. CAT expression from chimeric plasmids containing at least one copy of the HCMV 19 base pair (bp) repetitive motif was stimulated 10-fold in the presence of p13S-wt. The 19-bp motif contains a potential binding site for the cellular transcription factor ATF/CREB. Deletion analysis indicated that the ATF/CREB site was crucial for E1A-mediated stimulation. Insertion of a synthetic oligonucleotide homologous to a 19-bp motif and containing an ATF/CREB binding site into an HCMV chimera lacking ATF/CREB motifs conferred E1A responsivity on HCMV promoter-mediated CAT expression whereas insertion of a similar oligonucleotide containing a change of two bases in the sequence of the ATF/CREB site did not. Measurement of CAT-specific RNA verified the results of the CAT enzyme experiments. The ATF/CREB motif may be a target for stimulation of HCMV gene expression through either viral or cellular transcription factors.
Olive
D M
DM
Department of Microbiology, Faculty of Medicine, Kuwait University.
al-Mulla
W
W
Simsek
M
M
Zarban
S
S
al-Nakib
W
W
D051696:Activating Transcription Factors; D017456:Adenovirus Early Proteins; D000956:Antigens, Viral / Q000235:genetics*; D001483:Base Sequence; D001798:Blood Proteins / Q000378:metabolism; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002872:Chromosome Deletion; D017362:Cyclic AMP Response Element-Binding Protein; D004279:DNA, Viral / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D006367:HeLa Cells; D006801:Humans; D017874:Immediate-Early Proteins; D009841:Oligonucleotides / Q000096:biosynthesis / Q000235:genetics; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012367:RNA, Viral / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection
D051696:Activating Transcription Factors; D017456:Adenovirus Early Proteins; D000956:Antigens, Viral; D001798:Blood Proteins; D017362:Cyclic AMP Response Element-Binding Protein; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D017874:Immediate-Early Proteins; D009841:Oligonucleotides; D009856:Oncogene Proteins, Viral; D012367:RNA, Viral; D014157:Transcription Factors; C044582:immediate-early proteins, cytomegalovirus; D015500:Chloramphenicol O-Acetyltransferase