In humans, a polymorphic gene encodes the drug-metabolizing enzyme NAT1 (arylamine N-acetyltransferase Type 1), which is widely expressed throughout the body. While the protein-coding region of NAT1 is contained within a single exon, examination of the human EST (expressed sequence tag) database at the NCBI revealed the presence of nine separate exons, eight of which were located in the 5′ non-coding region of NAT1. Differential splicing produced at least eight unique mRNA isoforms that could be grouped according to the location of the first exon, which suggested that NAT1 expression occurs from three alternative promoters. Using RT (reverse transcriptase)-PCR, we identified one major transcript in various epithelial cells derived from different tissues. In contrast, multiple transcripts were observed in blood-derived cell lines (CEM, THP-1 and Jurkat), with a novel variant, not identified in the EST database, found in CEM cells only. The major splice variant increased gene expression 9–11-fold in a luciferase reporter assay, while the other isoforms were similar or slightly greater than the control. We examined the upstream region of the most active splice variant in a promoter-reporter assay, and isolated a 257 bp sequence that produced maximal promoter activity. This sequence lacked a TATA box, but contained a consensus Sp1 site and a CAAT box, as well as several other putative transcription-factor-binding sites. Cell-specific expression of the different NAT1 transcripts may contribute to the variation in NAT1 activity in vivo.
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Research Article|
March 22 2005
Genomic organization of human arylamine N-acetyltransferase Type I reveals alternative promoters that generate different 5′-UTR splice variants with altered translational activities
Neville J. BUTCHER;
Neville J. BUTCHER
1
1Centre for Medical Research, University of Western Australia, Nedlands, WA 6009, Australia
2Laboratory for Cancer Medicine, Western Australian Institute for Medical Research, Royal Perth Hospital, Perth, WA 6000, Australia
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Ajanthy ARULPRAGASAM;
Ajanthy ARULPRAGASAM
1Centre for Medical Research, University of Western Australia, Nedlands, WA 6009, Australia
2Laboratory for Cancer Medicine, Western Australian Institute for Medical Research, Royal Perth Hospital, Perth, WA 6000, Australia
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Hui Li GOH;
Hui Li GOH
1
1Centre for Medical Research, University of Western Australia, Nedlands, WA 6009, Australia
2Laboratory for Cancer Medicine, Western Australian Institute for Medical Research, Royal Perth Hospital, Perth, WA 6000, Australia
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Tamara DAVEY;
Tamara DAVEY
1Centre for Medical Research, University of Western Australia, Nedlands, WA 6009, Australia
2Laboratory for Cancer Medicine, Western Australian Institute for Medical Research, Royal Perth Hospital, Perth, WA 6000, Australia
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Rodney F. MINCHIN
1Centre for Medical Research, University of Western Australia, Nedlands, WA 6009, Australia
2Laboratory for Cancer Medicine, Western Australian Institute for Medical Research, Royal Perth Hospital, Perth, WA 6000, Australia
2To whom correspondence should be addressed, at the present address: School of Biomedical Sciences, University of Queensland, St. Lucia, QLD 4072, Australia (email r.minchin@uq.edu.au).
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Publisher: Portland Press Ltd
Received:
May 28 2004
Revision Received:
October 11 2004
Accepted:
October 18 2004
Accepted Manuscript online:
October 18 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2005
Biochem J (2005) 387 (1): 119–127.
Article history
Received:
May 28 2004
Revision Received:
October 11 2004
Accepted:
October 18 2004
Accepted Manuscript online:
October 18 2004
Citation
Neville J. BUTCHER, Ajanthy ARULPRAGASAM, Hui Li GOH, Tamara DAVEY, Rodney F. MINCHIN; Genomic organization of human arylamine N-acetyltransferase Type I reveals alternative promoters that generate different 5′-UTR splice variants with altered translational activities. Biochem J 1 April 2005; 387 (1): 119–127. doi: https://doi.org/10.1042/BJ20040903
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