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

被引:58
作者
Hsieh, Tsai-Yuan
Shiu, Tzu-Yue
Huang, Shih-Ming
Lin, Hsuan-Hwai
Lee, Tai-Chi
Chen, Peng-Jen
Chu, Heng-Cheng
Chang, Wei-Kuo
Jeng, King-Song
Lai, Michael M. C.
Chao, You-Chen
机构
[1] Natl Def Med Ctr, Dept Biochem, Taipei 114, Taiwan
[2] Tri Serv Gen Hosp, Dept Internal Med, Div Gastroenterol, Taipei, Taiwan
[3] Acad Sinica, Inst Mol Biol, Taipei 115, Taiwan
关键词
electrophoretic mobility shift assay; Gilbert's syndrome; TATA-binding protein; UDP-glucuronosyltransferase; 1A1;
D O I
10.1097/FPC.0b013e328012d0da
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Objectives Gilbert's syndrome is a congenital, nonhemolytic, unconjugated hyperbilirubinemia. The most common genotype of Gilbert's syndrome is the homozygous polymorphism, A(TA)(7)TAA, 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)(6)TAA or the mutant A(TA)(7)TAA 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-bincling 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)(5)TAA and A(TA)(8)TAA 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.
引用
收藏
页码:229 / 236
页数:8
相关论文
共 45 条
  • [1] ANALYSIS OF GENES FOR BILIRUBIN UDP-GLUCURONOSYLTRANSFERASE IN GILBERTS-SYNDROME
    AONO, S
    ADACHI, Y
    UYAMA, E
    YAMADA, Y
    KEINO, H
    NANNO, T
    KOIWAI, O
    SATO, H
    [J]. LANCET, 1995, 345 (8955): : 958 - 959
  • [2] Interleukin-1β inhibits CAR-induced expression of hepatic genes involved in drug and bilirubin clearance
    Assenat, E
    Gerbal-Chaloin, S
    Larrey, D
    Saric, J
    Fabre, JM
    Maurel, P
    Vilarem, MJ
    Pascussi, JM
    [J]. HEPATOLOGY, 2004, 40 (04) : 951 - 960
  • [3] Racial variability in the UDP-glucuronosyltransferase 1 (UGT1A1) promoter:: A balanced polymorphism for regulation of bilirubin metabolism?
    Beutler, E
    Gelbart, T
    Demina, A
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (14) : 8170 - 8174
  • [4] Biondi ML, 1999, CLIN CHEM, V45, P897
  • [5] HEPATIC BILIRUBIN UDP-GLUCURONYL TRANSFERASE ACTIVITY IN LIVER DISEASE AND GILBERTS SYNDROME
    BLACK, M
    BILLING, BH
    [J]. NEW ENGLAND JOURNAL OF MEDICINE, 1969, 280 (23) : 1266 - &
  • [6] Molecular diagnosis of a familial nonhemolytic hyperbilirubinemia (Gilbert's syndrome) in healthy subjects
    Borlak, J
    Thum, T
    Landt, O
    Erb, K
    Hermann, R
    [J]. HEPATOLOGY, 2000, 32 (04) : 792 - 795
  • [7] BOSMA PJ, 1994, J BIOL CHEM, V269, P17960
  • [8] THE GENETIC-BASIS OF THE REDUCED EXPRESSION OF BILIRUBIN UDP-GLUCURONOSYLTRANSFERASE-1 IN GILBERTS-SYNDROME
    BOSMA, PJ
    CHOWDHURY, JR
    BAKKER, C
    GANTLA, S
    DEBOER, A
    OOSTRA, BA
    LINDHOUT, D
    TYTGAT, GNJ
    JANSEN, PLM
    ELFERINK, RPJO
    CHOWDHURY, NR
    [J]. NEW ENGLAND JOURNAL OF MEDICINE, 1995, 333 (18) : 1171 - 1175
  • [9] WEIGHT MATRIX DESCRIPTIONS OF 4 EUKARYOTIC RNA POLYMERASE-II PROMOTER ELEMENTS DERIVED FROM 502 UNRELATED PROMOTER SEQUENCES
    BUCHER, P
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1990, 212 (04) : 563 - 578
  • [10] Tissue-specific, inducible, and hormonal control of the human UDP-glucuronosyltransferase-1 (UGT1) locus
    Chen, SJ
    Beaton, D
    Nguyen, N
    Senekeo-Effenberger, K
    Brace-Sinnokrak, E
    Argikar, U
    Remmel, RP
    Trottier, J
    Barbier, O
    Ritter, JK
    Tukey, RH
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (45) : 37547 - 37557