Thyroid Hormone Receptor Isoform-Specific Actions

甲状腺激素受体亚型特异性作用

• 批准号: 9222005
• 负责人: GREGORY A BRENT
• 金额: $ 33.5万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9222005
• 关键词: Adipocytes; Adult; Amino Acids; Amphibia; Binding; Binding Sites; Biological Assay; Biological Metamorphosis; Body Composition; Body Weight; Body Weight decreased; Catecholamines; Cell physiology; Cholesterol; Cholesterol Homeostasis; Consensus; Development; Dominant-Negative Mutation; Dyslipidemias; Energy Metabolism; Family; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Growth; Hydrophobicity; Infusion procedures; Lead; Ligands; Liver X Receptor; Lysine; Mammals; Mediating; Metabolic; Metabolic Diseases; Metabolic Pathway; Metabolism; Mitochondria; Mus; Mutant Strains Mice; Mutation; Nuclear; Nuclear Hormone Receptors; Nuclear Receptors; Obesity; PPAR alpha; Pathway interactions; Pattern; Peroxisome Proliferator-Activated Receptors; Pharmacology; Phenotype; Play; Post-Translational Protein Processing; Protein Isoforms; Proteins; Recruitment Activity; Regulation; Reporting; Repression; Respiration; Rest; Role; Signal Pathway; Signal Transduction; Site; Specificity; Thermogenesis; Thyroid Gland; Thyroid Hormone Receptor; Thyroid Hormone Receptor Gene; Thyroid Hormones; Thyroid hormone receptor alpha; Tissues; Transfection; Triiodothyronine; Ubiquitin; adipocyte differentiation; chromatin immunoprecipitation; cofactor; gene induction; gene repression; genome-wide; hormone regulation; in vitro Model; knock-down; lipid biosynthesis; metabolic rate; mouse model; mutant; novel; perilipin; public health relevance; relating to nervous system; response; therapeutic target; transcription factor; ubiquitin-protein ligase; uncoupling protein 1

DESCRIPTION (provided by applicant): Thyroid hormone is essential for normal development, growth, neural differentiation, and metabolic regulation in mammals, and is required for amphibian metamorphosis. There are two thyroid hormone receptor (TR) genes, TRα and TRβ, with different patterns of expression in development and in adult tissues. The relevance of TR isoform specificity is strongly supported by the distinct phenotypes reported in families heterozygous for dominant negative mutations of the TRβ gene and TRα gene. The mechanism of TR isoform-specific action, however, is not well understood. Posttranslational modification of nuclear receptors is being increasingly recognized as an important mechanism of gene regulation. Most proteins modified by small ubiquitin-like modifier (SUMO) are transcription factors and nuclear hormone receptors. Sumoylation requires conjugation of SUMO to a lysine within the consensus recognition motif ψ-Lys-X-Glu (ψ is a large hydrophobic amino acid) of a protein, which rapidly and dynamically modifies proteins involved in cellular processes. We determined that posttranslational modification of TR by conjugation of SUMO to TRα at lysines 283 and 389, and TRβ at lysines 50, 146 and 443, plays an essential role in triiodothyronine (T3)-induced gene induction and repression as well as TR isoform-specificity. TRα-SUMO conjugation is ligand-independent and utilizes the E3 ligase PIASxb. TRβ-SUMO is ligand-dependent and utilizes predominantly the E3 ligase PIAS1. SUMO1 and SUMO3 conjugation to TR are required for T3-dependent gene regulation, as demonstrated in transient transfection assay and studies of endogenous gene regulation. The role of SUMO1 and SUMO3 in T3-induction in transient functional assays is closely matched to the pattern of TR and co-factor binding in regulation of endogenous genes, as determined by Chromatin Immunoprecipitation (ChIP) assays. Co-repressors play an essential role in thyroid hormone action, and TR sumoylation is important for co-repressor recruitment. We have demonstrated that TR sumoylation is important for thyroid hormone-regulated metabolic actions including adipocyte differentiation and cross-talk with important metabolic regulators including LXR and PPARα. Specific aims that will be pursued include; 1. Utilize in vitro models to determine the role of TR sumoylation and TRα/TRβ isoform-specific actions in metabolic regulation. 2. Determine how sumoylation modulates TR-mediated gene regulation including TR interaction with transcription co-factors, metabolic signaling pathways, and recognition and binding to T3-regulated genes. 3. Evaluate the influence of TRα and TRβ sumoylation site mutations on thyroid hormone regulation of metabolism in a mouse model. We will assess the impact of introducing TRα (K389Q) and TRβ (K443Q) gene sumoylation mutations on the thyroid hormone axis, tissue level thyroid status, and metabolic regulation. Understanding the role of TR sumoylation in metabolic regulation and TR-isoform specific action may lead to the identification of novel thyroid hormone signaling targets relevant to therapies for metabolic diseases, including dyslipidemia and obesity.

描述(申请人提供)：甲状腺激素对哺乳动物的正常发育、生长、神经分化和代谢调节是必不可少的，也是两栖动物变态所必需的。甲状腺激素受体有两个基因，即甲状腺激素受体α和甲状腺激素受体β，它们在发育过程中和在成体组织中的表达模式不同。在trβ基因和trα基因的显性负突变杂合子家系中报告的不同表型有力地支持了tr异构体特异性的相关性。然而，tr亚型特异性作用的机制还不是很清楚。核受体的翻译后修饰正日益被认为是基因调控的一种重要机制。大多数被小泛素样修饰物(SUMO)修饰的蛋白质是转录因子和核激素受体。SUMO需要将SUMO连接到蛋白质的共识识别基序ψ-Lys-X-Glu(ψ是一种大的疏水氨基酸)中的赖氨酸，从而快速、动态地修饰参与细胞过程的蛋白质。我们确定，在三碘甲腺原氨酸(T3)诱导的基因诱导和抑制以及TRR的异构体特异性中，TR283和389位的SUMO与TRα以及50、146和443位的TRRβ结合的翻译后修饰在基因诱导和抑制中起着重要作用。TRα-SUMO结合是不依赖配体的，并利用E3连接酶PIASxb。TRβ-SUMO依赖于配体，主要利用E3连接酶PIAS1。瞬时基因转染法和内源性基因调控研究表明，依赖于T3的基因调控需要SUMO1和SUMO3结合到tr。在瞬时功能分析中，SUMO1和SUMO3在T3诱导中的作用与染色质免疫沉淀(ChIP)分析确定的内源基因调节中的TR和辅助因子结合模式密切匹配。辅抑制子在甲状腺激素的作用中起着至关重要的作用，而tr甲基化对于辅阻遏子的招募是重要的。我们已经证明，在甲状腺激素调节的代谢活动中，包括脂肪细胞分化和与重要的代谢调节因子，包括LXR和PPARα的相互作用，TRSUMO化是重要的。将追求的具体目标包括：1.利用体外模型来确定tr苏莫化和trα/trβ异构体特异性作用在代谢调节中的作用。2.确定苏莫化是如何调节tr介导的基因调控的，包括tr与转录辅助因子的相互作用，代谢信号通路，以及对t3调节基因的识别和结合。3.在小鼠甲状腺激素代谢调节模型中，评价了trα和trβ苏莫化位点突变对甲状腺激素代谢调节的影响。我们将评估引入trα(K389Q)和trβ(K443Q)基因总合突变对甲状腺激素轴、组织水平甲状腺状态和代谢调节的影响。了解tr总甲基化在代谢调节中的作用和tr异构体的特异性作用可能有助于识别与治疗代谢疾病相关的新的甲状腺激素信号靶点，包括血脂异常和肥胖症。

项目成果

Persistent COUP-TFII expression underlies the myopathy and impaired muscle regeneration observed in resistance to thyroid hormone-alpha.

• DOI: 10.1038/s41598-021-84080-5
• 发表时间: 2021-02-25
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Aguiari P;Liu YY;Petrosyan A;Cheng SY;Brent GA;Perin L;Milanesi A
• 通讯作者: Milanesi A

Beam Me In: Thyroid Hormone Analog Targets Alternative Transporter in Mouse Model of X-Linked Adrenoleukodystrophy.

向我发送信息：甲状腺激素类似物靶向 X 连锁肾上腺脑白质营养不良小鼠模型中的替代转运蛋白。

• DOI: 10.1210/en.2017-00206
• 发表时间: 2017
• 期刊: Endocrinology
• 影响因子: 4.8
• 作者: Milanesi,Anna;Brent,GregoryA
• 通讯作者: Brent,GregoryA

Thyroid hormone receptor sumoylation is required for preadipocyte differentiation and proliferation.

甲状腺激素受体苏酰化是前脂肪细胞分化和增殖所必需的。

• DOI: 10.1074/jbc.m114.600312
• 发表时间: 2015
• 期刊: The Journal of biological chemistry
• 作者: Liu,Yan-Yun;Ayers,Stephen;Milanesi,Anna;Teng,Xiaochun;Rabi,Sina;Akiba,Ysutada;Brent,GregoryA
• 通讯作者: Brent,GregoryA