Analysis of thyroxine-binding globulin (TBG) gene regulation and inherited TGB abnormalities

甲状腺素结合球蛋白 (TBG) 基因调控和遗传性 TGB 异常分析

• 批准号: 04671468
• 负责人: MORI Yuichi
• 金额: $ 1.34万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04671468/
• 关键词: Thyroxine-binding globulin; Complete deficiency; Partial deficiency; Excess; Frameshift mutation; Gene screening; Intracellular transport; HNF-1; TBG異常症; 遺伝子解析; クローニング; 転写調節

1. Complete TGB deficiency in Japanese(TBG-CDJ)has a nucleotide deletion at codon 352 among 395 resulting in the production of C-terminal 22 amino acid truncation due to a frameshift and premature termination. The cDNA of TBG-CDJ was expressed in COS-1 cells. Pulse chase experiment revealed TBG-CDJ was retained in the rough endoplasmic reticulum resulting in complete deficiency in serum.Gene screening showed 23 Japanese families with TBG-CD had a same mutation TBG-CDJ.2. Abnormality of X-chromosome inactivation was found in a female TBG-CD patient. She was heterozygous of TBG-CDJ and normal TBG alleles. The latter allele was selectively inactivated.3. Partial TBG deficiency in Japanese (TBG-PDJ) has a nucleotide substitution at codon 363 replacing normal pro (CCT) with leu (CTT). The TBG-PDJ was expressed in COS-1 cells. lt was revealed that delay of intracellular transport of TBG-PDJ was a cause of partial deficiency. Induction of GRP78, molecular chaperone, was found to participate the transport disorder.4. Gene duplication was shown in a hereditary TBG excess family by using PCR and HPLC.5. Human and rat genomic TBG genes were isolated and characterized. Promoter regions of both genes were sequenced and subcloned to CAT or luciferase reporter construct. HNF-1 (hepatocyte nuclear factor 1) binding site was found important fot basal and liver specific expression. Gene expression and serum. TBG were down regulated by corticosterone in rat.

1.日本人完全性TGB缺乏症（TBG-CDJ）在395个密码子中的352个处具有核苷酸缺失，导致由于移码和过早终止而产生C-末端22个氨基酸截短。TBG-CDJ基因在COS-1细胞中得到表达。结论：1.脉冲追踪实验显示TBG-CDJ滞留在粗面内质网中，导致血清中完全缺失，基因筛查显示23个日本TBG-CD家系存在相同的TBG-CDJ突变。1例女性TBG-CD患者X染色体失活异常。她是TBG-CDJ和正常TBG等位基因杂合子。后一个等位基因被选择性失活.日本人的部分TBG缺乏症（TBG-PDJ）在密码子363处有一个核苷酸取代，将正常的pro（CCT）替换为leu（CTT）。TBG-PDJ在COS-1细胞中表达。提示TBG-PDJ细胞内转运延迟是部分缺乏的原因。发现分子伴侣GRP 78的诱导参与了转运障碍. PCR和HPLC检测结果表明，该遗传性TBG过多家系存在基因重复.人和大鼠基因组TBG基因的分离和表征。对两个基因的启动子区进行测序并亚克隆到CAT或荧光素酶报告基因构建体中。肝细胞核因子1（hepatocyte nuclear factor 1，HNF-1）结合位点对基础表达和肝脏特异性表达有重要作用。基因表达和血清。皮质酮可下调大鼠TBG。

项目成果

Charles H.Emerson: "Gene expression and serum thyroxine-binding globulin are regulated by adrenal status and corticosterone in the rat." Endocrinology. 133. 1192-1196 (1993)

Charles H.Emerson：“基因表达和血清甲状腺素结合球蛋白受大鼠肾上腺状态和皮质酮的调节。”

Yamamori Ikuo: "A nucleotide deletion is universal as a cause of complete thyroxine-binding globulin deficiency in Japanese." The 9th International Congress of Endocrinology,Abstract. 05-01-128 (1992)

Yamamori Ikuo：“核苷酸缺失是日本人甲状腺素结合球蛋白完全缺乏的普遍原因。”

Kyoto Takeda et al.: "Gene screening in Japanese families with thyroxine binding globulin complete deficiency demonstrates that a nucleotide deletion at codon 352 may be a rase specific mutation" Clin Endocrinol. (in press).

京都武田 (Kyoto Takeda) 等人：“对甲状腺素结合球蛋白完全缺乏的日本家庭进行的基因筛查表明，密码子 352 处的核苷酸缺失可能是一种酶特异性突变”Clin Endocrinol。

Fukushi Kambe: "An additional carbohydrate chain in the variant thyroxine-binding globulin(TBG asp-96)impairs its secretion" Molecular Endocrinology. 6. 443-449 (1992)

Fukushi Kambe：“变体甲状腺素结合球蛋白 (TBG asp-96) 中的额外碳水化合物链会损害其分泌”《分子内分泌学》。

Tsukasa Noguchi et al.: " Hereditary complete thyroxine-binding globulin deficiency : identification by T3 resin uptake test and DNA analysis" Internal medicine. 32. 6-9 (1993)

Tsukasa Noguchi 等人：“遗传性完全甲状腺素结合球蛋白缺乏症：通过 T3 树脂摄取试验和 DNA 分析进行鉴定”内科。