title: Coordinated regulation of transcription and alternative splicing by the thyroid hormone receptor and its associating coregulators.
abstract: Emerging evidence has indicated that the transcription and processing of precursor mRNA (pre-mRNA) are functionally coupled to modulate gene expression. In collaboration with coregulators, several steroid hormone receptors have previously been shown to directly affect alternative pre-mRNA splicing coupled to hormone-induced gene transcription; however, the roles of the thyroid hormone receptor (TR) and its coregulators in alternative splicing coordinated with transcription remain unknown. In the present study, we constructed a luciferase reporter and CD44 alternative splicing (AS) minigene driven by a minimal promoter carrying 2 copies of the palindromic thyroid hormone-response element. We then examined whether TR could modulate pre-mRNA processing coupled to triiodothyronine (T3)-induced gene transcription using luciferase reporter and splicing minigene assays in HeLa cells. In the presence of cotransfected TRβ1, T3 increased luciferase activities along with the inclusion of the CD44 variable exons 4 and 5 in a dose- and time-dependent manner. In contrast, cotransfected TRβ1 did not affect the exon-inclusion of the CD44 minigene driven by the cytomegalovirus promoter. T3-induced two-exon inclusion was significantly increased by the cotransfection of the TR-associated protein, 150-kDa, a subunit of the TRAP/Mediator complex that has recently been shown to function as a splicing factor. In contrast, T3-induced two-exon inclusion was significantly decreased by cotransfection of the polypyrimidine tract-binding protein-associated splicing factor, which was previously shown to function as a corepressor of TR. These results demonstrated that liganded TR in cooperation with its associating cofactors could modulate alternative pre-mRNA splicing coupled to gene transcription.
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan. Electronic address: tsato@gunma-u.ac.jp.
authors.forename: Tetsurou
authors.initials: T
authors.lastname: Satoh
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Katano-Toki
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Tomaru
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yoshino
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Ishizuka
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Horiguchi
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Yasuyo
authors.initials: Y
authors.lastname: Nakajima
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Sumiyasu
authors.initials: S
authors.lastname: Ishii
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ozawa
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Shibusawa
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Koshi
authors.initials: K
authors.lastname: Hashimoto
authors.affiliation: Kitakanto Molecular Novel Research Institute for Obesity and Metabolism, Midori, Japan.
authors.forename: Masatomo
authors.initials: M
authors.lastname: Mori
authors.affiliation: Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Masanobu
authors.initials: M
authors.lastname: Yamada
mesh_terms: D017398:Alternative Splicing; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005091:Exons; D006367:HeLa Cells / Q000187:drug effects; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D000072670:PTB-Associated Splicing Factor; D011401:Promoter Regions, Genetic; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D011988:Receptors, Thyroid Hormone / Q000235:genetics* / Q000378:metabolism; D020218:Response Elements; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014284:Triiodothyronine / Q000494:pharmacology
chemical_list: C497483:CD44 protein, human; D004268:DNA-Binding Proteins; D018960:Hyaluronan Receptors; D000072670:PTB-Associated Splicing Factor; D016601:RNA-Binding Proteins; D011988:Receptors, Thyroid Hormone; C475774:THRAP3 protein, human; D014157:Transcription Factors; D014284:Triiodothyronine; D008156:Luciferases
keywords: 150-kDa; Alternative splicing; Polypyrimidine tract-binding protein-associated splicing factor; Thyroid hormone receptor; Thyroid hormone receptor-associated protein; Transcription