Characterization of the 5′-flanking region of the human RNA-specific adenosine deaminase ADAR1 gene and identification of an interferon-inducible ADAR1 promoter

The double-stranded RNA-specific adenosine deaminase (ADAR1) is inducible by interferon (IFN) and is implicated in the editing of viral RNAs during lytic and persistent infection. We have now isolated and characterized human genomic clones that contain the promoter region required for transcription...

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Veröffentlicht in:Gene 1999-03, Vol.229 (1), p.203-213
Hauptverfasser: George, C.X., Samuel, C.E.
Format: Artikel
Sprache:eng
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Zusammenfassung:The double-stranded RNA-specific adenosine deaminase (ADAR1) is inducible by interferon (IFN) and is implicated in the editing of viral RNAs during lytic and persistent infection. We have now isolated and characterized human genomic clones that contain the promoter region required for transcription of the ADAR1 gene. Rapid amplification of cDNA 5′-ends (5′-RACE) identified additional upstream exon 1 sequence that was localized on P1-phage and λ-phage genomic clones by Southern gel-blot analysis and sequence analysis. A Northern gel-blot analysis using a probe corresponding to the 5′-RACE exon 1 sequence and adjacent exon 2 sequence detected a major RNA transcript of ∼6.7 kb that was IFN-inducible in human amnion U cells. Transient transfection assays, using chloramphenicol acetyltransferase (CAT) as the reporter in constructs possessing various 5′-flanking fragments of the ADAR1 gene, led to the identification of a functional TATA-less promoter that directed IFN-inducible transcription of CAT. Sequence determination and deletion analysis of the promoter region revealed a consensus copy of the I FN- S timulated R esponse E lement (ISRE) involved in IFN inducibility that was flanked by a K inase C onserved S equence (KCS)-like element previously found to be unique to the human and mouse PKR gene promoters. A 63-bp minimal promoter fragment possessing the KCS-like and ISRE elements was sufficient to drive IFN-inducible transcription.
ISSN:0378-1119
1879-0038
DOI:10.1016/S0378-1119(99)00017-7