DNA binding and interaction with the nuclear receptor corepressor of thyroid hormone receptor are required for ligand-independent stimulation of the mouse preprothyrotropin-releasing hormone gene

被引:25
作者
Satoh, T
Monden, T
Ishizuka, T
Mitsuhashi, T
Yamada, M
Mori, M
机构
[1] Gunma Univ, Sch Med, Dept Internal Med 1, Maebashi, Gumma 3718511, Japan
[2] Univ Tokyo, Fac Med, Dept Internal Med 3, Tokyo 113, Japan
关键词
corepressor; mouse TRH gene; negative regulation; thyroid hormone receptor;
D O I
10.1016/S0303-7207(99)00032-5
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
A negative thyroid hormone response element (TRE) in the mouse preprothyrotropin-releasing hormone (TRH) gene was previously mapped within the proximal promoter element between - 83 and + 53 that contained a TRE half-site motif at - 57 ((-57)TGACCT(-52)). In transfection experiments, the promoter activity is stimulated by unliganded thyroid hormone receptor (TR) and T-3 reverses the basal promoter stimulation. In this study, we determined whether the direct binding of TR to the TRE half-site in the mouse TRH gene is required for the ligand-independent stimulation using a transient transfection assay into CV-1 cells and electrophoretic mobility shift assays (EMSA). In addition, the role of a corepressor protein for the ligand-independent stimulation was examined using a putative splicing variant of the nuclear receptor corepressor (N-CoRI). Point mutations introduced into the TRE half-site at - 57 eliminated the binding of TR and the stimulatory effect of unliganded TR. Two mutant TRs lacking DNA-binding activity and two CoR box mutant TRs showed no stimulation in the wild-type TRH promoter. The cotransfected N-CoRI potentiated the ligand-independent stimulation by the wild-type TR, but did not compensate for the impaired function of the CoR box mutant TR. In EMSA, TR strongly bound as homodimers and weakly as heterodimers with retinoid X receptor (RXR) to the element containing the TRE half-site at - 57. Binding of TR to the TRE half-site was essential to form homo- and heterodimers, and the RXR binding site appeared to be located downstream of the TRE half-site. In vitro translated N-CoRI preferentially bound TR homodimers over TR/RXR heterodimers. These results collectively suggest that the DNA-bound TR/corepressor complex might be directly involved in the ligand-independent stimulation of the mouse TRH gene promoter. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.
引用
收藏
页码:137 / 149
页数:13
相关论文
共 55 条
  • [1] Role for N-CoR and histone deacetylase in Sin3-mediated transcriptional repression
    Alland, L
    Muhle, R
    Hou, H
    Potes, J
    Chin, L
    SchreiberAgus, N
    DePinho, RA
    [J]. NATURE, 1997, 387 (6628) : 49 - 55
  • [2] Ausubel FM., 1994, Curr. Protoc. Mol. Biol
  • [3] BODENNER DL, 1991, J BIOL CHEM, V266, P21666
  • [4] BURNSIDE J, 1989, J BIOL CHEM, V264, P6886
  • [5] At least three subdomains of v-erbA are involved in its silencing function
    Busch, K
    Martin, B
    Baniahmad, A
    Renkawitz, R
    Muller, M
    [J]. MOLECULAR ENDOCRINOLOGY, 1997, 11 (03) : 379 - 389
  • [6] THYROID-HORMONES REGULATE RAT THYROTROPIN-BETA GENE PROMOTER ACTIVITY EXPRESSED IN GH3 CELLS
    CARR, FE
    BURNSIDE, J
    CHIN, WW
    [J]. MOLECULAR ENDOCRINOLOGY, 1989, 3 (04) : 709 - 716
  • [7] NOMENCLATURE OF THYROID-HORMONE RECEPTOR BETA-GENE MUTATIONS IN RESISTANCE TO THYROID-HORMONE - CONSENSUS STATEMENT FROM THE FIRST WORKSHOP ON THYROID-HORMONE RESISTANCE, JULY 10-11, 1993, CAMBRIDGE, UNITED-KINGDOM
    CHATTERJEE, VKK
    BECKPECCOZ, P
    CHIN, WW
    DEGROOT, LJ
    JAMESON, JL
    NAKAMURA, H
    REFETOFF, S
    USALA, SJ
    WEINTRAUB, BD
    [J]. JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM, 1994, 78 (04) : 990 - 993
  • [8] SMRT isoforms mediate repression and anti-repression of nuclear receptor heterodimers
    Chen, JD
    Umesono, K
    Evans, RM
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (15) : 7567 - 7571
  • [9] A TRANSCRIPTIONAL CO-REPRESSOR THAT INTERACTS WITH NUCLEAR HORMONE RECEPTORS
    CHEN, JD
    EVANS, RM
    [J]. NATURE, 1995, 377 (6548) : 454 - 457
  • [10] CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2