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.

p130 cas是一种含有SH 3结构域和多个酪氨酸残基的对接蛋白。p130 cas位于粘着斑，对整联蛋白刺激作出反应而被酪氨酸磷酸化，并被认为通过c-Crk和其它蛋白质将信号传递给肌动蛋白应力纤维的重塑和细胞运动。在通过Far Western筛选寻找p130 cas的SH 3结构域配体的过程中，我们克隆了一个新的蛋白质CIZ，它由一个亮氨酸拉链、一个富含丝氨酸/苏氨酸的区域、一个富含脯氨酸的序列、5、6或8个Kruppel型C2 H2锌指和谷氨酰胺-丙氨酸重复序列组成。CIZ与细胞中的Cas结合，位于细胞核和粘着斑处。我们发现，CIZ是一个核质穿梭蛋白，使用瞬时种间异核体形成试验。为了寻找CIZ在细胞核中的靶点，我们通过CASTing分析确定CIZ的DNA结合一致性为（G/C）AAAAA（A）。基质金属蛋白酶是降解细胞外基质蛋白的酶，其启动子中存在类似共有序列。CIZ与MMP-1（胶原酶）启动子中的共识样序列结合。CIZ的过表达上调MMP-1、MMP-3（基质溶解素）和MMP-7（基质溶解素）启动子的转录，并且这种转录激活在Cas存在时增强。此外，CIZ的稳定过表达促进了培养基中MMP-7的产生。综上所述，CIZ是一种新型锌指蛋白，结合Cas，是一种核质穿梭蛋白，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。

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

Nakamoto T, Yamagata T, Sakai R, Ogawa S, Honda H, Ueno H, Hirano N, Yazaki Y, Hirai H.: "CIZ : a zinc finger protein that interacts with p130cas and activates the expression of matrix metalloproteinases."Mol.Cell.Biol.. 20. 1649-1658 (2000)

Nakamoto T、Yamagata T、Sakai R、Okawa S、Honda H、Ueno H、Hirano N、Yazaki Y、Hirai H.：“CIZ：一种与 p130cas 相互作用并激活基质金属蛋白酶表达的锌指蛋白。”Mol。

Honda H: "p130Cas, an assembling molecule of actin filaments, promotes cell movement, cell migration, and cell spreading in fibroblasts"Biochem. Biophys. Res. Commun.. 262. 25-30 (1999)

Honda H：“p130Cas，一种肌动蛋白丝的组装分子，促进成纤维细胞中的细胞运动、细胞迁移和细胞扩散”Biochem。