Anakysis of GATA-3 function in proliferation and differentiation of kerationocytes

GATA-3在角质细胞增殖和分化中的功能分析

• 批准号: 13670865
• 负责人: KAWACHI Yasuhiro
• 金额: $ 1.6万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670865/
• 关键词: GATA-3; Keratinocytes; 表皮; ロリクリン; keratinocytes

In epidermal keratinocytes, we demonstrated that GATA-3 was more abundant in differentiated keratinocytes than undifferentiated keratinocytes, suggesting that GATA-3 might contribute to differntiatioirspecific gene regulation. We previously showed that c-Fos and Sp1 activate the loricrin promoter activity in differentiated keratinocytes while c-Jun and Sp3 suppress it in undifferentiated keratinocytes. Thus, we tested the cooperative tarnsactivation of loricrin promoter activity by GATA-3, c-Fos and Sp1. Transactivation experiments presented here provide evidence that GATA-3, c-Fos and Sp1 can activate the loricrin promoter transcription in a synergistic manner even though the GATA-3 binding motif is deleted from the promoter. These results strongly suggest the interaction among those proteins. Thus, we examined the physical binding among GATA-3, AP-1 and Sp1/Sp3. In vitro binding by means of a glutathione S-transferase pull down assay showed that c-Jun/c-Fos could directly associate with GATA-3. However, binding affinity of GATA-3 with Sp1 was much less than that with c-Fos, These results suggest that GATA-3 can interact with c-Fos directly while GATA-3 may interact with Sp1 via other transcription factors or cofactors.

在表皮角质形成细胞中，我们发现GATA-3在分化的角质形成细胞中比未分化的角质形成细胞中更丰富，这表明GATA-3可能参与分化特异性基因调控。我们之前发现c-Fos和Sp1在分化的角质形成细胞中激活loricrin启动子活性，而c-Jun和Sp3在未分化的角质形成细胞中抑制它。因此，我们测试了GATA-3、c-Fos和Sp1对氯丙林启动子活性的协同作用。本研究的反式激活实验证明，即使GATA-3结合基序从启动子中删除，GATA-3、c-Fos和Sp1也能以协同方式激活氯丙氨酸启动子转录。这些结果有力地说明了这些蛋白质之间的相互作用。因此，我们研究了GATA-3、AP-1和Sp1/Sp3之间的物理结合。体外结合谷胱甘肽s -转移酶拉下实验表明，c-Jun/c-Fos可直接与GATA-3结合。然而，GATA-3与Sp1的结合亲和力远低于与c-Fos的结合亲和力，这些结果表明GATA-3可以直接与c-Fos相互作用，而GATA-3可能通过其他转录因子或辅因子与Sp1相互作用。

项目成果

Waikel RL, Kawachi Y et al.: "Deregulation expression of c-Myc depletes epidermal stem cells"Nature Genetics. 28. 165-168 (2001)

Waikel RL、Kawachi Y 等人：“c-Myc 表达失调会消耗表皮干细胞”《自然遗传学》。

Kotsuji-Maruyama T, Imakado S, Kawachi Y, Otsuka F.: "PDGF-BB induces MAP kinase phosphorylation and VEGF expression in neurofibroma-derived cultured cells from patients with neurofibromatosis-1"Journal of Dermatology. 29. 713-717 (2002)

Kotsuji-Maruyama T、Imakado S、Kawachi Y、Otsuka F.：“PDGF-BB 在神经纤维瘤病患者的神经纤维瘤衍生培养细胞中诱导 MAP 激酶磷酸化和 VEGF 表达 - 1”皮肤病学杂志。

Waikel RL., Kawachi Y. et al.: "Deregulated expression of c-Myc depletes epidermal sterm cells"Nature Genetics. 28(2). 165-168 (2001)

Waikel RL.、Kawachi Y. 等人：“c-Myc 表达失调会消耗表皮干细胞”《自然遗传学》。

Kawachi Y et al.: "Expression of angiogenic factors in neurofibromas"Experimental Dermatology. 11. 1-6 (2003)

Kawachi Y 等人：“神经纤维瘤中血管生成因子的表达”实验皮肤病学。

Kotsuji T.Kawachi Y et al.: "PDGF-BB induces MAP kinase phosphorylation and VEGF expression in neurofibroma-derived cultured cells from patients with NF-1"Journal of Dermatology. 29. 713-717 (2002)

Kotsuji T.Kawachi Y 等人：“PDGF-BB 在来自 NF-1 患者的神经纤维瘤衍生培养细胞中诱导 MAP 激酶磷酸化和 VEGF 表达”《皮肤病学杂志》。