Screening of novel signal transducers for BMP using DNA affinity purification

使用 DNA 亲和纯化筛选新型 BMP 信号转导器

• 批准号: 15591611
• 负责人: IMAMURA Takeshi
• 金额: $ 2.24万
• 依托单位: Japanese Foundation for Cancer Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591611/
• 关键词: BMP; Smad; Gcn5; 蛋白質-DNA相互作用; シグナル伝達

Members of the transforming growth factor-β (TGF-β) superfamily, including TGF-β, activin, nodal, and bone morphogenetic proteins (BMPs), are multifunctional cytokines that regulate a wide range of cellular responses, including cell proliferation, differentiation, adhesion, migration, and apoptosis. TGF-β and related proteins transduce signals through two distinct serine/threonine kinase receptors, termed type I and type II, and intracellular Smad proteins. Eight different Smad proteins have been identified in mammals, and are classified into three groups : receptor-regulated Smads (R-Smads), common-partner Smad (Co-Smad), and inhibitory Smads (I-Smads). Smad2 and Smad3 are R-Smads activated by TGF-β/activin/nodal receptors ALK-4, -5, and -7, whereas Smad1, Smad5, and Smad8 are BMP-specific R-Smads. Smad4 is the Co-Smad shared by signaling pathways for TGF-β and activin and those for BMPs. Smad6 and Smad7 are I-Smads in mammals ; Smad6 preferentially suppresses BMP signaling, whereas S … More mad7 inhibits both BMP and TGF-β signaling.The roles of TGF-β in cancer biology are complex ; TGF-β can suppress or promote tumor growth depending on the type of cancer. The ability of TGF-β to potently inhibit the proliferation of epithelial, endothelial, and hematopoietic cell lineages is central to its tumor-suppressive effects. However, as tumors evolve, they often become refractory to TGF-β-mediated growth inhibition and overexpress TGF-β, which induces epithelial-to-mesenchymal transition (EMT) of tumor cells and facilitates immunosuppression, extracellular matrix deposition, and angiogenesis. It was recently reported that inhibition of autocrine TGF-β signaling in carcinoma cells reduces cell invasiveness and tumor metastasis, and that these effects of TGF-β are closely associated with the ability of TGF-β to induce EMT and stimulate cell migration. The TGF-β signaling pathway has correspondingly become an attractive target for drug development in the field of oncology.To identify new components of transcriptional complexes containing Smad proteins, we purified DNA-binding proteins from human breast cancer MCF-7 cell nuclear extract using a Smad-binding DNA element as bait, and identified a co-activator GCN5 as a direct partner of activated Smad complexes. GCN5 is structurally similar to PCAF, which was previously identified as a co-activator for R-Smads for TGF-β signaling pathways. GCN5 functions like PCAF, in that it binds to TGF-β-specific R-Smads, and enhances transcriptional activity induced by TGF-β. In addition, GCN5, but not PCAF, interacts with R-Smads for BMP signaling pathways, and enhances BMP-induced transcriptional activity, suggesting that GCN5 and PCAF have distinct physiological functions in vivo. Moreover, silencing of the GCN5 gene by RNA interference results in repression of transcriptional activities induced by TGF-β. Less

转化生长因子-β（TGF-β）超家族的成员，包括TGF-β、激活素、结节和骨形态发生蛋白（BMP），是调节广泛的细胞反应的多功能细胞因子，包括细胞增殖、分化、粘附、迁移和凋亡。TGF-β和相关蛋白通过两种不同的丝氨酸/苏氨酸激酶受体（称为I型和II型）和细胞内Smad蛋白进行信号传导。哺乳动物中已经鉴定出8种不同的Smad蛋白，并将其分为三组：受体调节的Smad（R-Smad）、共同伴侣Smad（Co-Smad）和抑制性Smad（I-Smad）。Smad 2和Smad 3是由TGF-β/激活素/nodal受体ALK-4、ALK-5和ALK-7激活的R-Smads，而Smad 1、Smad 5和Smad 8是BMP特异性R-Smads。Smad 4是TGF-β和激活素信号通路以及BMPs信号通路共享的Co-Smad。Smad 6和Smad 7是哺乳动物中的I-Smads; Smad 6优先抑制BMP信号传导，而S ...更多信息 mad 7抑制BMP和TGF-β信号传导。TGF-β在癌症生物学中的作用是复杂的; TGF-β可以根据癌症的类型抑制或促进肿瘤生长。TGF-β有效抑制上皮、内皮和造血细胞谱系增殖的能力是其肿瘤抑制作用的核心。然而，随着肿瘤的发展，它们通常变得对TGF-β介导的生长抑制不敏感并过表达TGF-β，这诱导肿瘤细胞的上皮向间充质转化（EMT）并促进免疫抑制、细胞外基质沉积和血管生成。最近报道，抑制癌细胞中自分泌的TGF-β信号传导降低细胞侵袭和肿瘤转移，并且TGF-β的这些作用与TGF-β诱导EMT和刺激细胞迁移的能力密切相关。为了鉴定含有Smad蛋白的转录复合物的新组分，我们以Smad结合DNA元件为诱饵，从人乳腺癌MCF-7细胞核提取物中纯化了DNA结合蛋白，并鉴定了一个共激活因子GCN 5作为激活的Smad复合物的直接配偶体。GCN 5在结构上类似于PCAF，PCAF先前被鉴定为TGF-β信号通路的R-Smads的共激活剂。GCN 5的功能类似于PCAF，因为它结合TGF-β特异性R-Smads，并增强TGF-β诱导的转录活性。此外，GCN 5，而不是PCAF，与BMP信号通路的R-Smads相互作用，并增强BMP诱导的转录活性，表明GCN 5和PCAF在体内具有不同的生理功能。此外，通过RNA干扰沉默GCN 5基因导致TGF-β诱导的转录活性的抑制。少

项目成果

Coordinate regulation of cell growth and differentiation by TGF-β superfamily and Runx proteins

• DOI: 10.1038/sj.onc.1207131
• 发表时间: 2004-05-24
• 期刊: ONCOGENE
• 影响因子: 8
• 作者: Miyazono, K;Maeda, S;Imamura, T
• 通讯作者: Imamura, T

Roles for the MH2 domain of Smad7 in the specific inhibition of transforming growth factor-β superfamily signaling

• DOI: 10.1074/jbc.m313977200
• 发表时间: 2004-07-23
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Mochizuki, T;Miyazaki, H;Miyazono, K
• 通讯作者: Miyazono, K

Kahata K, Imamura T (外7名): "Regulation of transforming growth factor-b and bone morphogenetic protein signaling by transcriptional co-activator GCN5"Genes Cells. 9・2. 143-151 (2004)

Kahata K、Imamura T（其他 7 人）：“转录共激活因子 GCN5 对转化生长因子-b 和骨形态发生蛋白信号的调节”Genes Cells 9·2。

Miyazono K, Suzuki H, Imamura T: "Regulation of TGF-b signaling and its roles in progression of tumors."Cancer Sci. 94・3. 230-234 (2003)

Miyazono K、Suzuki H、Imamura T：“TGF-b 信号传导及其在肿瘤进展中的作用”。Cancer Sci. 94・3 (2003)。

Regulation of transforming growth factor-β and bone morphogenetic protein signaling by transcriptional co-activator GCN5.

转录共激活因子 GCN5 对转化生长因子-β 和骨形态发生蛋白信号的调节。

• 发表时间: 2004
• 期刊: Genes Cells. 9
• 作者: Kahata K;Hayashi M;Asaka M;Hellman U;Kitagawa H;Yanagisawa J;Kato S;Imamura T;Miyazono M
• 通讯作者: Miyazono M