Analyses of Maltipotential Functions of a Novel Signaling Molecule, Cas

新型信号分子 Cas 的多电位功能分析

• 批准号: 11694250
• 负责人: HIRAI Hisamaru
• 金额: $ 5.57万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A).
• 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-11694250/
• 关键词: Cas; CIZ; focal adhesion; SH3 domain; matrix metalloproteinase; actin stress fiber; nucleo-cytoplasmic shuttling protein; インテグリン; SH3; マトリックスメタロプロテアーゼ; プロモーター; 転写制御

p130cas is a docking protein that contains an SH3 domain and multiple tyrosine residues. p130cas is located at focal adhesions, is tyrosine phosphorylated in response to integrin stimulation, and is thought to transmit signals, via c-Crk and other proteins, to the remodeling of actin stress fibers and to the cell movement. In the search for the ligands of the SH3 domain of p130cas by Far Western screening, we cloned a novel protein named CIZ.CIZ consists of a putative leucine zipper, serine/threonine rich region, a proline rich sequence, 5, 6 or 8 Kruppel-type C2H2 zinc fingers, and the glutamine-alanine repeat. CIZ binds Cas in cells and is located in nucleus and at focal adhesions. We showed that CIZ is a nucleo-cytoplasmic shuttling protein, using the transient interspecies heterokaryon formation assay. In order to search for the targets of CIZ in nucleus, we determined the DNA binding consensus of CIZ as (G/C)AAAAA(A) by CASTing analysis. The consensus-like sequences are found in several promoters of matrix metalloproteinases, which are the enzymes to degrade the extracellular matrix proteins. CIZ binds to a consensus-like sequence in MMP-1 (collagenase) promoter. Overexpression of CIZ upregulates the transcriptions from MMP-1, MMP-3 (stromelysin), and MMP-7 (matrilysin) promoters, and this transactivation was enhanced in the presense of Cas. Furthermore, the stable overexpression of CIZ promoted the production of MMP-7 in culture medium. In summary, CIZ, a novel zinc finger protein, binds Cas, is a nucleo-cytoplasmic shuttling protein, and regulates the expression of matrix metalloproteinases.

P130CAS是一种对接蛋白，其中包含SH3结构域和多个酪氨酸残基。 P130CAS位于局灶性粘附处，是对整联蛋白刺激的酪氨酸磷酸化的，被认为可以通过C-CRK和其他蛋白质将信号传输到肌动蛋白应激纤维的重塑和细胞运动。在搜索P130CAS的SH3域的配体时，我们通过远西方筛选了，我们克隆了一种名为Ciz的新型蛋白-Type C2H2锌指的手指，谷氨酰胺 - 丙氨酸重复。 CIZ在细胞中结合Cas，位于核中和局灶性粘连中。我们表明CIZ是一种使用瞬态杂质杂质者形成测定法，是一种核胞质穿梭蛋白。为了在核中搜索CIZ的靶标，我们通过铸造分析确定了CIZ AS（g/c）AAAAA（a）的DNA结合共识。在基质金属蛋白酶的几个启动子中发现了类似共识的序列，这些酶是降解细胞外基质蛋白的酶。 CIZ与MMP-1（胶原酶）启动子中的共有序列结合。 CIZ的过表达上调了MMP-1，MMP-3（Stromelysin）和MMP-7（母氏菌蛋白）启动子的转录，并且在CAS的标志中增强了这种反式激活。此外，CIZ的稳定过表达促进了培养基中MMP-7的产生。总而言之，CIZ是一种新型的锌指蛋白，结合Cas，是一种核总质式穿梭蛋白，并调节基质金属蛋白酶的表达。

项目成果

Nakamoto T: "CIZ : a zinc finger protein that interacts with p130cas and activates the expression of matrix metalloproteinases"Mol. Cell. Biol.. (In press).

Nakamoto T：“CIZ：一种锌指蛋白，与 p130cas 相互作用并激活基质金属蛋白酶的表达”Mol。

Honda H: "Acquired loss of p53 induced blastic transformation of p210bcr/abl-expressing hematopoietic cells : A mouse model for blastic crisis of human CML"Blood. (In press).

Honda H：“获得性 p53 缺失诱导 p210bcr/abl 表达造血细胞的急变转化：人类 CML 急变危机的小鼠模型”血液。

Furuya K: "Overexpression of Cas-interacting zinc finger protein (CIZ) suppresses proliferation and enhances expression of Type I collagen gene in osteoblast-like MC3T3El cells."Exp.Cell Res.. 261. 329-335 (2001)

Furuya K：“Cas 相互作用的锌指蛋白 (CIZ) 的过度表达抑制增殖并增强成骨细胞样 MC3T3E1 细胞中 I 型胶原基因的表达。”Exp.Cell Res.. 261. 329-335 (2001)

Yamagata T: "Acetylation of GATA-3 affects T-cell survival and homing to secondary lymphoid organs."EMBO J.. 19. 4676-4687 (2000)

Yamagata T：“GATA-3 的乙酰化影响 T 细胞存活和归巢至次级淋巴器官。”EMBO J.. 19. 4676-4687 (2000)

Kanda H, Mimura T, Hamasaki K, Yamamoto K, Yazaki Y, Hirai H, Nojima Y.Fyn and Lck tyrosine kinases: "regulate tyrosine phosphorylation of p105CasL, a member of the p130Cas docking protein family, in T-cell receptor-mediated signaling."Immunol.. 97. 56-61

Kanda H、Mimura T、Hamasaki K、Yamamoto K、Yazaki Y、Hirai H、Nojima Y.Fyn 和 Lck 酪氨酸激酶：“在 T 细胞受体介导中调节 p105CasL（p130Cas 对接蛋白家族成员）的酪氨酸磷酸化