アデノウイルズDNAの複製開始領域中の転写プロモーター活性
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概要
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The replication origin of adenovirus DNA resides within the terminal 51 base pair(bp) sequences (nucleotides 1 to 51) at either terminus of a linear viral genome. The terminal 18 bp sequences (nucleotides 1 to 18) are absolutely required for initiation reactions and are referred to as the minimal origin. The minimal origin contains a perfectly conserved AT-rich sequence (5' GGTATATTAT 3', nucleotides 9 to 18) in human adenoviruses which resembles TATA box sequences of promoters from higher eukaryote genes. The next 33bp sequences (nucleotides 19 to 51) efficiently increase the replication activity of the minimal origin by interacting with cellular transcription factors; nuclear factor I/CCAAT-binding transcription factor (NFI/CTF) and nuclear factor III/octamer transcription factor-1 (NFIII/OTF-1). We examined whether the adenovirus replication origin had transcriptional promoter activity. We constructed test plasmids containing adenovirus left-terminal fragments inserted upstream of the promoter-removed neomycin resistant gene (neo gene) which confers G418 (neomycin-analogue)-resistant phenotype on mammalian cells. After transfections of test plasmids into cells, transient expression and long term expression of the neo gene were determined by S1 nuclease analysis of cytoplasmic RNA. The expression level of the neo gene was also estimated by counting G418-resistant cell colonies. The results showed that the neo gene was transcribed in cells transfected with the test plasmids containing adenovirus left-terminal fragments, but not in cells transfected with the test plasmid containing no exogenous DNA sequences. Initiation sites of the neo gene transcripts were located approximately 30bp downstream from the AT sequence of the minimal origin, suggesting that the minimal origin has a TATA boxlike function. The results suggest that the adenovirus replication origin is a potential promoter of transcription.
- 札幌医科大学の論文
- 1989-10-01
札幌医科大学 | 論文
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