Abstract
Human UDP-glucuronosyltransferase (UGT) is a part of a major excretion pathway for endobiotics and xenobiotics. The UGT family of genes is highly polymorphic, and our aim is to describe novel polymorphisms at the UGT1A3 locus and determine how they alter substrate metabolism and drug reactions. One hundred healthy Japanese adults volunteered for the present study. We sequenced PCR-amplified fragments of the gene directly, and calculated the frequency of the genetic variations detected. To measure variant enzyme activity, we constructed five expression models and used estrone as the substrate in the assays. We identified six novel single nucleotide polymorphisms (SNPs). Of these, four caused amino acid substitutions (17A→G: Q6R, 31T→C: W11R, 133C→T: R45W, and 140T→C: V47A) and the remaining two were silent (81G→A: E27E and 447A→G: A159A). We found five types of alleles having differing SNP combinations: wild type (frequency=0.61), W11R-E27E-A159A (0.10), Q6A-W11R-E27E-A159A (0.055), W11R-E27E-V47A-A159A (0.125), and R45W (0.11). Expression studies found that the variants changed the enzyme efficiencies (K m/V max) to 121% of the wild type for W11R, 86% for Q6R-W11R, 369% for W11R-V47A, and 70% for R45W. Several UGT 1A3 polymorphisms exist in the Japanese population, having different levels of activity. These polymorphisms are capable of affecting the steady state levels of estrogens, and may increase sensitivity to adverse drug effects.
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Acknowledgements
We thank Prof. K. Horiike of the Department of Biochemistry I, Shiga University of Medical Science, for helpful suggestions and for comments on the kinetic analysis of UGT1A3. We also thank Mr M. Suzaki of the Central Research Laboratory, Shiga University of Medical Science, for technical assistance. This work was partly supported by grants-in-aid for scientific research from the Ministry of Education, Science, and Culture of Japan (14704032 and 12204061) and the Morinaga Hoshi-kai (Tokyo, Japan).
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The accession numbers of new nucleotides: AB120359, AB120360, AB120361, AB120362.
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Iwai, M., Maruo, Y., Ito, M. et al. Six novel UDP-glucuronosyltransferase (UGT1A3) polymorphisms with varying activity. J Hum Genet 49, 123–128 (2004). https://doi.org/10.1007/s10038-003-0119-y
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DOI: https://doi.org/10.1007/s10038-003-0119-y
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