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Quantitation and characterization of N‐acetyltransferase‐2 mRNA in human tissues
Author(s) -
Husain Anwar,
Barker David F,
Zhang Xiaoyan,
Doll Mark A,
States J. Christopher,
Hein David W.
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1180-b
N‐acetyltransferase 2 (NAT2) mRNA was examined by 5′‐RLM‐RACE and review of public database information to more precisely localize the NAT2 promoter(s) and to validate the design of a quantitative RT‐PCR assay. The great majority (40/41) of NAT2 cDNAs prepared by 5′‐RLM‐RACE of liver RNA had 5′ termini between 8682 and 8752 nucleotides upstream of the NAT2 ORF exon, and 34/40 of these 5′‐termini were at the 8711 and 8716 adenines. All of 59 NAT2 cDNAs with 5′ termini in this vicinity, including 40 5′‐RACE cDNAs from liver and 19 cDNAs in public databases from liver and other sources, showed direct splicing to the ORF exon, with no other non‐coding exon detected. A sensitive and specific TaqMan RT‐PCR assay with intron‐spanning primers was developed and used, together with a second TaqMan RT‐PCR assay based on amplification of a NAT2 ORF exon segment, to measure NAT2 mRNA in 29 different human tissues. The close correlation of the mRNA measurements obtained with the intron‐spanning primer assay and the intra‐ORF primer assay indicate a single major NAT2 promoter. NAT2 mRNA was highest in liver, small intestine and colon and readily detected in most other tissues albeit at levels 1% or less than in liver. Expression of very low levels of NAT2 mRNA in various human tissues may be relevant to associations between NAT2 genotype and arylamine‐induced cancers at these tumor sites. Partially supported by USPHS grants CA34627 & ES12557.