Structural Organization and Transcriptional Regulation of the Human beta_1-Adrenergic Receptor Gene

人类β_1-肾上腺素能受体基因的结构组织和转录调控

• 批准号: 01440048
• 负责人: MATSUI Hiroaki
• 金额: $ 16万
• 依托单位: ST. MARIANNA UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01440048/
• 关键词: beta_1-adrenergic receptor; antidepressant; receptor regulation; receptor gene expression; βーアドレナリン受容体; 受容体遺伝子発現調節; 抗うつ薬作用機序; βアドレナリン受容体; 受容体遺伝子発現調節機構

To investigate the effects of antidepressant treatment on the beta_1-adrenergic receptor(AR) at transcriptional levels, the delineation of the structural organization of the human beta_1-AR gene was attempted. A genomic clone was obtained from human placental genomic library through repeated screenings using the 1.3-kb(kilobase) Smal fragment human beta_1-AR cDNA(complementary DNA) probe. Restriction endonuclease fragments of interest were subcloned into pUC18 and sequenced by the dideoxy chain-termination method.The proximal promoter region (-1300 through -50 relative to the adenosine of the first methionine codon) lacks TATA box, is rich in G + C content, and has multiple putative binding sites for transcriptional factor Sp1 and AP-2. Thus, the proximal promoter region has features of "house- keeping" genes.The distal promoter region (-2300 through -1300) has representative TATA box and CAAT box consequences, typical thyroid, estrogen, glucorticoid, cAMP responsive el'ements, and binding sites for transcriptional factor AP-1, GATA-1,2.3, which indicate multiple regulatory mechanisms involved in the beta_1-AR gene expression in vivo. Moreover, there is present in the human genome a long [1032-bp(base pair)] open reading frame which ends only 112-bp 5' to the initiator methionine of the receptor, and which encodes a putative protein of the molecular weight of - 38000.

为了研究抗抑郁治疗在转录水平上对 β_1-肾上腺素能受体 (AR) 的影响，尝试描绘人类 β_1-AR 基因的结构组织。使用1.3-kb（千碱基）Smal片段人β_1-AR cDNA（互补DNA）探针反复筛选，从人胎盘基因组文库中获得基因组克隆。将感兴趣的限制性内切酶片段亚克隆到pUC18中，并通过双脱氧链终止法进行测序。近端启动子区（相对于第一个蛋氨酸密码子的腺苷-1300到-50）缺乏TATA盒，富含G + C含量，并且具有转录因子 Sp1 和 AP-2 的多个假定结合位点。因此，近端启动子区域具有“管家”基因的特征。远端启动子区域（-2300至-1300）具有代表性的TATA盒和CAAT盒结果，典型的甲状腺、雌激素、糖皮质激素、cAMP反应元件，以及转录因子 AP-1、GATA-1,2.3 的结合位点，表明体内 beta_1-AR 基因表达涉及多种调控机制。此外，人类基因组中存在一个长的[1032-bp（碱基对）]开放阅读框，其末端仅位于受体起始子蛋氨酸的5'端112-bp，并且编码分子量为- 38000。

项目成果

朝倉 幹雄,: "抗うつ薬投与後のラット脳内存性蛋白質のCa^<2+>/リン脂質依存性蛋白質リン酸化." 精神薬療基金研究年報,. 22,. 163-168 (1991)

Mikio Asakura，：“服用抗抑郁药后内源性大鼠脑蛋白的 Ca^2+/磷脂依赖性蛋白磷酸化。”精神药理学治疗基金年度报告，22, 163-168 (1991)

Kenichi Osada: "Effects of phorbol ester on the membrane-inserted protein kinase C." Japanese J. Psychopharmacology. 10. 204 (1990)

Kenichi Osada：“佛波酯对膜插入蛋白激酶 C 的影响。”

Hiroaki Matsui: "Effects of the chronic antidepressant treatment on the transcriptional regulation of the betaー1 adrenergic receptor gene." FEBS Letters.

Hiroaki Matsui：“长期抗抑郁治疗对 β-1 肾上腺素受体基因转录调节的影响。”

松井 宏晃: "神経生化学" (日本生化学会編,東京化学同人), 10 (1990)

松井宏明：《神经生物化学》（日本生物化学会编、东京化学同人），10（1990）

Atsushi Adachi: "Evidence for the involvement of guanine nucleotide binding proteins in the regulation of α_1Aーand α_1Bーadrenergic receptors in the rat cerebral cortex." European Neuropsychopharmacology.

Atsushi Adachi：“鸟嘌呤核苷酸结合蛋白参与调节大鼠大脑皮层 α_1A 和 α_1B 肾上腺素受体的证据。”