Transcriptional coactivator UTF1 as an embryonic stem cell cofactor for retinoic acid receptor

转录辅激活因子 UTF1 作为视黄酸受体的胚胎干细胞辅因子

• 批准号: 11670133
• 负责人: OKUDA Akihiko
• 金额: $ 2.3万
• 依托单位: Saitama Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670133/
• 关键词: mammal; early embryogenesis; embryonic carcinoma; retinoic acid; transcriptional cofactor; 哺乳動物初期胚; ES細胞; コアクティベーター

Embryonic carcinoma (EC) cell lines such as F9 and P19 cells are widely used as a model system for studying the molecular basis governing early developmental stages of mammals. The EC cells possess a number of properties which resemble those of early embryonic cells. For example, these EC cells retain pluripotent properties and can be induced to differentiate into several types of cells such as muscle and nerve cells using retinoic acid and/or by aggregate formation. Early studies carried out in EC cells clearly indicated that these cells have a number of interesting and unique regulatory features. One of such examples is that about the regulation of retinoic acid receptor (RAR) β2 gene expression. The promoter of this gene carries two RAR responsive element (RARE) and its activity is autoregulated by its own gene. However, it has been demonstrated that the gene requires an additional embryonic stem cell cofactor for its expression..In 1998, I have molecularly cloned a novel transcript … More ional coactivator UTF1 whose expression is restricted to early embryonic cells. When I looked the amino acid sequence carefully, I found that the protein contains two LxxLL motif in the one of the evolutionarily conserved domain which is commonly present in nuclear receptor cofactor such as SRC-1 and TIF-2 and is demonstrated to be involved in the interaction with nuclear receptor. Therefore, I hypothesized that the UTF1 may correspond to the embyonic stem cell specific cofactor required for RARβ2 gene expression. By the transient transfection, I demonstrated that UTF1 is able to stimulate transcription from this promoter. Furthermore, I found that this activation of transcription is occurred through the RARE.I also demonstrated, by GST pull down assay, that UTF1 is able to bind to RAR directly. However, this interaction is found to be independent of retinoic acid and AF2 domain of the RAR.Thus, the UTF1 appears to bind to RAR in a distinct way from other nuclear receptor cofactors such as SRC-1 and TIF-2. Less

胚胎癌（EC）细胞系（例如F9和P19细胞）被广泛用作研究哺乳动物早期发育阶段的分子基础的模型系统。 EC细胞具有许多类似于早期胚胎细胞的特性。例如，这些EC细胞保留多能特性，并可以通过视黄酸和/或通过骨料形成分化为几种类型的细胞，例如肌肉和神经细胞。在EC细胞中进行的早期研究清楚地表明，这些细胞具有许多有趣且独特的调节特征。这样的例子之一是关于视黄酸受体（RAR）β2基因表达的调节。该基因的启动子具有两个RAR响应元件（稀有），其活性由其自身基因自动调节。然而，已经证明该基因需要一个额外的胚胎干细胞辅因子的表达。.在1998年，我分子克隆了一个新型的转录本……更具效率共激活器UTF1的表达仅限于早期胚胎细胞。当我仔细看氨基酸序列时，我发现该蛋白在一个进化构成的结构域中包含两个LXXLL基序，该结构域通常存在于核接收器辅助因子中，例如SRC-1和TIF-2，并且被证明与核接收器相互作用。因此，我假设UTF1可能对应于RARβ2基因表达所需的Embyonic干细胞特异性辅因子。通过瞬态翻译，我证明了UTF1能够刺激该启动子的转录。此外，我发现这种转录的激活是通过稀有发生的。我还通过GST下拉分析证明了UTF1能够直接结合RAR。但是，发现这种相互作用与雷尔的视黄酸和AF2结构域无关。较少的

项目成果

Ogawa, S., Fujita, M., Ishii, Y., Tsurukami, H., Hirabayashi, M., Ikeda, K., Orimo, A., Hosoi, T., Ueda, M., Nakamura, T., Ouchi, Y., Muramatsu, M., and Inoue, S.: "Impaired estrogen sensitivity in bone by inhibiting both estrogen receptor alpha and beta

小川，S.，藤田，M.，石井，Y.，鹤上，H.，平林，M.，池田，K.，奥里莫，A.，细井，T.，上田，M.，中村，T.，

Orimo, A., Inoue, S., Minowa, O., Tominaga, N., Sato, M., Kuno, J., Hiroi, H., Shimizu, Y., Suzuki, M., Noda, T., and Muramatsu, M.: "Undeveloped uterus and reduced estrogen responsiveness in mice with dsruption of the estrogen-responsive finger protein g

奥里莫，A.，井上，S.，箕轮，O.，富永，N.，佐藤，M.，久野，J.，广井，H.，清水，Y.，铃木，M.，野田，T.，

Akira. Orimo: "Underdeveloped uterus and reduced estrogen responsiveness in mice with disruption of the estrogen-responsive finger protein gene, which is a direct target of estrogen receptor α"Proc. Natl. Acad. Sci.. 96. 12027-12032 (1999)

Akira. Orimo：“雌激素反应性指蛋白基因被破坏，导致小鼠子宫发育不全，雌激素反应性降低，该基因是雌激素受体 α 的直接靶点”Proc.. 12027-12032（1999 年）。 ）

Akiko Fukushima: "Carboxy-terminally truncated form of a coactivator UTF1 stimulates transcription from a variety of gene promoters through the TATA box"Biochem.Biophys.Res.Comun.. 258. 519-523 (1999)

Akiko Fukushima：“共激活子 UTF1 的羧基末端截短形式通过 TATA 盒刺激多种基因启动子的转录”Biochem.Biophys.Res.Comun.. 258. 519-523 (1999)

Fukushima, A., Nishimoto, M., Okuda, A.and Muramatsu, M.: "Carboxy-terminally truncated form of a coactivator UTF1 stimulates transcription from a variety of gene promoters through the TATA box."Biochem.Biopys.Res.Commun.. 258. 519-523 (1999)

Fukushima, A.、Nishimoto, M.、Okuda, A. 和 Muramatsu, M.：“共激活子 UTF1 的羧基末端截短形式通过 TATA 盒刺激多种基因启动子的转录。”Biochem.Biopys.Res。