Molecular pathogenesis of Gilbert's syndrome: Decreased TATA-binding protein binding affinity of UGT1A1 gene promoter

Tsai-Yuan Hsieh, Tzu-Yue Shiu, Shih-Ming Huang, Hsuan-Hwai Lin, Tai C. Lee, Peng-Jen Chen, Heng-Cheng Chu, W.-K. Chang, King-Song Jeng, Michael M.C. Lai, Y.-C. Chao

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44 Citations (Scopus)

Abstract

OBJECTIVES: Gilbert's syndrome is a congenital, nonhemolytic, unconjugated hyperbilirubinemia. The most common genotype of Gilbert's syndrome is the homozygous polymorphism, A(TA)7TAA, in the promoter of the gene for UDP-glucuronosyltransferase 1A1 (UGT1A1), with a thymine adenine insertion in the TATA-box-like sequence, which results in a decrease in UGT1A1 activity. The mechanism responsible for this decrease in UGT1A1 activity, however, has not been elucidated. To clarify the mechanism underlying this deficiency in UGT1A1 activity in patients with Gilbert's syndrome. METHODS: The promoter activity assay using the wild-type A(TA)6TAA or the mutant A(TA)7TAA promoter and a luciferase reporter was performed in two different hepatoma cell lines. The binding affinity for a nuclear protein complex or for TATA-binding protein was evaluated by a competitive electophoretic mobility shift assay using wild-type or mutant TATA-box-like oligonucleotide probes and nuclear extract or TATA-binding protein. The formation of complexes between TATA-binding protein and wild-type or mutant oligonucleotide probes was also studied by a quantitive electophoretic mobility shift assay. RESULTS: A TA insertion in the TATA-box-like sequence of the promoter activity of UGT1A1 gene. A competitive electrophoretic mobility shift assay showed a decrease in nuclear proteincomplex binding affinity and TATA-binding protein binding affinity of the mutant TATA-box-like sequence A(TA)7TAA. When the mutants A(TA)5TAA and A(TA)8TAA were also compared, quantitative electrophoretic mobility shift assay demonstrated that the TATA-binding protein binding affinity progressively decreased as the number of TA repeats in the TATA-box-like sequence increased. CONCLUSION: TA insertion in the TATA-box-like sequence of the UGT1A1 promoter affected its binding affinity for TATA-binding protein, causing a decrease in its activity. This explains the pathogenesis of Gilbert's syndrome. © 2007 Lippincott Williams & Wilkins, Inc.
Original languageEnglish
Pages (from-to)229-236
Number of pages8
JournalPharmacogenetics and Genomics
Volume17
Issue number4
DOIs
Publication statusPublished - 2007
Externally publishedYes

Keywords

  • Electrophoretic mobility shift assay
  • Gilbert's syndrome
  • TATA-binding protein
  • UDP-glucuronosyltransferase 1A1
  • glucuronosyltransferase 1A1
  • oligonucleotide
  • TATA binding protein
  • amino acid sequence
  • article
  • binding affinity
  • controlled study
  • enzyme activity
  • gel mobility shift assay
  • gene activity
  • gene insertion
  • Gilbert disease
  • hepatoma cell
  • human
  • human cell
  • oligonucleotide probe
  • pathogenesis
  • priority journal
  • promoter region
  • protein binding
  • Base Sequence
  • Cell Line
  • DNA Primers
  • Electrophoretic Mobility Shift Assay
  • Gilbert Disease
  • Glucuronosyltransferase
  • Humans
  • Kinetics
  • Mutagenesis, Insertional
  • Pharmacogenetics
  • Promoter Regions (Genetics)
  • TATA-Box Binding Protein

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    Hsieh, T-Y., Shiu, T-Y., Huang, S-M., Lin, H-H., Lee, T. C., Chen, P-J., Chu, H-C., Chang, W-K., Jeng, K-S., Lai, M. M. C., & Chao, Y-C. (2007). Molecular pathogenesis of Gilbert's syndrome: Decreased TATA-binding protein binding affinity of UGT1A1 gene promoter. Pharmacogenetics and Genomics, 17(4), 229-236. https://doi.org/10.1097/FPC.0b013e328012d0da