Structure and Functional Analysis of Bone Morphogenetic Protein Receptor

骨形态发生蛋白受体的结构与功能分析

• 批准号: 06454592
• 负责人: UENO Naoto
• 金额: $ 4.29万
• 依托单位: HOKKAIDO UNIEVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454592/
• 关键词: BMP; bone formation; receptor; Ser; Thr kinase; スレオニンキナーゼ; 骨形成因子; TGF-βスーパーファミリー; 自己リン酸化

We have isolated several novel receptor Ser/Thr kinase cDNAs from a mouse MC3T3-E1 cells cDNA using oligonucleotide primers designed in the conserved sequence among TGF-beta family receoptors such as activin and TGF-beta type II receptors. One of them designated as mTFR11turned out to be a type I class of receptor that binds both BMP-2 and BMP-4. We constructed a mutantreceptor cDNA that predicts a truncated receptor lacking entire Ser/Thr kinase and confirmed that the dominant negative receptor inhibited both BMP-2 and BMP-4 in cultured osteoblastic cells and in early embryos. Moreover, we generated constitutively active form ofthe BMP receptor by a point mutation Gln to Asp in GS domain, which is believed to be essential for BMP signaling. We further confirmed that the mutant receptor transmits BMP-like signals independent of BMP ligands.Our effort is now focused on the identification of downstream factor that mediates BMP signaling. Therefore, we have screened a human cDNA library by yeast two-hybrid method using a cytoplasmic region of the BMP receptor as a bait. Successfully, we have isolated three cDNAs that encodes proteins that binds BMP receptor and function of the proteins in the signal transduction is currently investigated.

我们利用在TGF-β家族受体如激活素和TGF-β II型受体的保守序列中设计的寡核苷酸引物，从小鼠MC 3 T3-E1细胞cDNA中分离了几个新的受体Ser/Thr激酶cDNA。其中一个被命名为mTFR 11的受体被证明是I型受体，它结合BMP-2和BMP-4。我们构建了一个mu受体cDNA，预测一个截短的受体缺乏整个丝氨酸/苏氨酸激酶，并证实，显性负性受体抑制BMP-2和BMP-4在培养的成骨细胞和早期胚胎。此外，我们通过GS结构域中Gln到Asp的点突变产生了BMP受体的组成型活性形式，这被认为是BMP信号传导所必需的。我们进一步证实了该突变体受体不依赖BMP配体而传递BMP样信号。因此，我们利用BMP受体的胞浆区作为诱饵，通过酵母双杂交方法筛选了人cDNA文库。我们成功地分离出了三个编码BMP受体结合蛋白的cDNA，目前正在研究这些蛋白在信号转导中的功能。

项目成果

Yamaguchi et al.: "Identification of a member of the MAPKKK family as a potrntial mediator of THF-β signal transduction" Science. 270. 2008-2011 (1995)

Yamaguchi 等人：“鉴定 MAPKKK 家族成员作为 THF-β 信号转导的潜在介质”《科学》270。2008-2011 (1995)

Mishima et al.: "Bmpr encodes a type I bone morphpgenetic protein receptor that is essential for gastmlation during mpuse embryogenisis" Genes & Dev.9. 3027-3037 (1995)

Mishima 等人：“Bmpr 编码一种 I 型骨形态发生蛋白受体，该受体对于 mpuse 胚胎发生过程中的胃化至关重要”

鈴木厚: "「アフリカツメガエルの背腹軸形成」Annual Review細胞生物学" 中外医学社, 9 (1994)

Atsushi Suzuki：“‘爪蟾背腹轴形成’细胞生物学年度评论”Chugai Igakusha，9 (1994)

Suzuki, A., Shioka, N., Maeda, J., Tada, M.and Ueno, N: "Cloning of an isoform of mouse TGF-b type II receptor gene" FEBS Letters. 355. 19-22 (1994)

Suzuki, A.、Shioka, N.、Maeda, J.、Tada, M. 和 Ueno, N：“小鼠 TGF-b II 型受体基因亚型的克隆”FEBS Letters。

A.Suzuki: "A truncated BMP receptor affects dorsal-ventral patterning in the early Xenopus embryo." Proc.Natl.Acad.Sci.USA.91. 10255-10259 (1994)

A.Suzuki：“截短的 BMP 受体影响早期非洲爪蟾胚胎的背腹模式。”