Drug Metabolism and Pharmacokinetics
Online ISSN : 1880-0920
Print ISSN : 1347-4367
ISSN-L : 1347-4367
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An Artifact Derived from a Pseudogene Led to the Discovery of MicroRNA Binding Site Polymorphism in the 3′-Untranslated Region of the Human Dihydrofolate Reductase Gene
Hideki HAYASHIYui TAZOEMasato HORINOChihiro FUJIMAKI-KATOHSeiji TSUBOITaiji MATSUYAMAKazuhiro KOSUGEHiroshi YAMADADaiki TSUJIKazuyuki INOUEKunihiko ITOH
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2012 Volume 27 Issue 2 Pages 263-267

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Abstract

  A novel single-nucleotide polymorphism (SNP) in the 3′-untranslated region of the human dihydrofolate reductase (DHFR) gene with enhanced expression was identified in 2001. In 2007, it was reported that this SNP, DHFR C829T, was located close to a microRNA binding site and contributed to the stability of mRNA. Many researchers have analyzed this SNP in several races including Asians and Caucasians. However, the mutation allele is not yet confirmed in most populations. In this study, we reinvestigated the frequency of this SNP using three methods. First, this SNP in genomic DNA was analyzed by a PCR-restriction fragment length polymorphism method. Second, this SNP in mRNA was analyzed by a single nucleotide extension method following a reverse transcription reaction. Third, the mRNA expression level was analyzed by a real-time PCR method. The findings in our study, regarding the discovery of this SNP, suggest that the SNP is an artifact caused by contamination by the genomic DNA of the pseudogene DHFRP1. This study is a reinvestigation of a newly discovered genetic polymorphism.

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© 2012 by The Japanese Society for the Study of Xenobiotics
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