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，淋巴结和骨形态发生蛋白（BMP），是多功能细胞因子，可调节广泛的细胞反应，包括细胞增殖，分化，粘合剂，粘合剂，迁移和凋亡。 TGF-β和相关蛋白通过两个不同的丝氨酸/苏氨酸激酶受体转化信号，称为I型和II型和细胞内SMAD蛋白。在哺乳动物中已经鉴定出八种不同的SMAD蛋白，并分为三组：受体调节的SMAD（R-SMADS），普通伴侣SMAD（Co-SMAD）和抑制性SMADS（I-SMADS）。 SMAD2和SMAD3是由TGF-β/Activin/Nodal受体ALK-4，-5和-7激活的R-SMADS，而SMAD1，SMAD5和SMAD8是BMP特异性的R-SMADS。 SMAD4是TGF-β和激活素的信号通路以及BMP的信号通路共享的共有共同的SMAD。 smad6和smad7是哺乳动物中的i-smads； SMAD6优先抑制BMP信号传导，而S…更多的MAD7抑制了BMP和TGF-β信号传导。TGF-β在癌症生物学中的作用很复杂。 TGF-β可以根据癌症的类型抑制或促进肿瘤生长。 TGF-β潜在地抑制上皮，内皮和造血细胞谱系的增殖的能力对于其肿瘤抑制作用至关重要。然而，随着肿瘤的发展，它们通常会对TGF-β介导的生长抑制和过表达TGF-β的耐受性，这会诱导肿瘤细胞的上皮到间质转变（EMT），并促进免疫抑制，细胞外基质下降下降和血管生成。最近据报道，癌细胞中自分泌TGF-β信号传导的抑制可降低细胞的侵袭性和肿瘤转移，而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。 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在结构上与PCAF相似，PCAF先前被确定为用于TGF-β信号通路的R-SMADS的共激活因子。 GCN5像PCAF一样起作用，因为它与TGF-β特异性R-SMADS结合，并增强TGF-β诱导的转录活性。此外，GCN5而不是PCAF与R-SMAD相互作用，用于BMP信号通路，并增强BMP诱导的转录活性，这表明GCN5和PCAF在体内具有不同的生理功能。此外，通过RNA干扰对GCN5基因沉默会导致TGF-β诱导的转录活性的表达。较少的

项目成果

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)。

Matsuyama S, Imamura T (外9名): "B-431542 and Gleevec inhibit TGF-b induced growth stimulation human osteosarcoma cells."Cancer Res.. 63・22. 7791-7798 (2003)

Matsuyama S、Imamura T（其他 9 人）：“B-431542 和格列卫抑制 TGF-b 诱导的生长刺激人骨肉瘤细胞。”Cancer Res.. 63・22 (2003)。

SB-431542 and Gleevec inhibit TGF-β induced growth stimulation of human osteosarcoma cells.

SB-431542 和 Gleevec 抑制 TGF-β 诱导的人骨肉瘤细胞的生长刺激。

• 发表时间: 2003
• 期刊: Cancer Res. 63・22
• 作者: Matsuyama S;Imamura T (外9名)
• 通讯作者: Imamura T (外9名)