Signal transduction mechanisms through granulocyte colony-stimulating factor receptor.

通过粒细胞集落刺激因子受体的信号转导机制。

• 批准号: 09680638
• 负责人: MURAKAMI Hiroshi
• 金额: $ 2.05万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680638/
• 关键词: granulocyte; gene-expression; receptor; signal-transduction; proliferation; differentiation; tyrosine-kinase; colony-stimulating-factor

To analyze the mechanisms of neutrophil differentiation, murine homologue (MZF-2) of human MZF-1 cDNA, which encodes the transcription factor. involved in neutrophil differentiation, was isolated. C-terminus region of MZF-2 protein containing zinc finger domain is highly homologous to that of the MZF-1, However, the amino-terminus of MZF-2 protein was different from that of MZF-1 protein in that MZF-2 protein has leucine rich region (LeR domain) and a region rich in acidic amino acids. The chromosomal gene of MZF-2 consist of 5 exons and alternative transcription start sites of the gene produce MZF-1 and MZF-2 mRNAs. The analysis of transcription activation activities of amino-terminus deletion derivatives of MZF-2 revealed that amino-terminus region interacted to the transcription-activation domain and suppressed its own transcriptional activation.On the other hand, G-CSF dependent phospho-proteins, which can bind to the tyrosine-phosphorylated G-CSF receptor, were isolated. The N-terminal amino-acid sequence of one of those was identical to that of SHIP.Its G-CSF dependent tyrosine phosphorylation and interaction with G-CSF receptor were demonstrated.Furthermore, I examined G-CSF dependent expression of several anti-apoptotic genes. One of them, bcl-2, was found to be expressed about 3 hours after G-CSF stimulation. As the gene was not expressed in cells carrying G-CSF receptor with mutated 4th cytoplasmic tyrosine residue, the 4th tyrosine residue of the receptor seemed to be involved in the anti-apoptotic signal transduction.Moreover, alpha-D-mannnosidase and macrophage inflammatory protein 1-alpha genes were found to be expressed 15 minutes after G-CSF stimulation.

为了分析中性粒细胞分化的机制，人类 MZF-1 cDNA 的鼠同源物 (MZF-2) 编码转录因子。参与中性粒细胞分化，被分离出来。 MZF-2蛋白的C端区域含有锌指结构域，与MZF-1的C端区域高度同源，但MZF-2蛋白的氨基端与MZF-1蛋白的不同之处在于MZF-2蛋白具有富含亮氨酸区域（LeR结构域）和富含酸性氨基酸的区域。 MZF-2 的染色体基因由 5 个外显子组成，该基因的替代转录起始位点产生 MZF-1 和 MZF-2 mRNA。对MZF-2氨基末端缺失衍生物的转录激活活性分析表明，氨基末端区域与转录激活结构域相互作用并抑制其自身的转录激活。可以与酪氨酸磷酸化 G-CSF 受体结合，被分离出来。其中一个的 N 端氨基酸序列与 SHIP 相同。证明了其 G-CSF 依赖性酪氨酸磷酸化以及与 G-CSF 受体的相互作用。此外，我检查了几种抗凋亡药物的 G-CSF 依赖性表达。基因。其中之一，bcl-2，被发现在 G-CSF 刺激后约 3 小时表达。由于该基因在携带胞质第4酪氨酸残基突变的G-CSF受体的细胞中不表达，因此该受体的第4酪氨酸残基似乎参与了抗凋亡信号转导。此外，α-D-甘露糖苷酶和巨噬细胞炎症蛋白发现 G-CSF 刺激 15 分钟后 1-α 基因表达。

项目成果

Murai, K., Murakami, H., and Nagata, S: "Myeloid-specific transcriptional activation by murine myeloid zinc-finger protein 2." Proc.Natl.Acad.Sci.USA. 95. 3461-3466 (1998)

Murai, K.、Murakami, H. 和 Nagata, S：“小鼠骨髓锌指蛋白 2 的骨髓特异性转录激活。”

A. Thomson: "The cytokine handbook, the third edition." Academic press., 927 (1998)

A. Thomson：“细胞因子手册，第三版。”

Orita, T.: "Binding of NF-Y transcription factor to one of the cis-elements in the myeloperoxidase gene promoter that responds to granulocyte colony-stimulating factor." J.Biol.Chem.272. 23216-23223 (1997)

Orita, T.：“NF-Y 转录因子与响应粒细胞集落刺激因子的髓过氧化物酶基因启动子中的顺式元件之一结合。”

Murai, K., Murakami, H., and Nagata, S.: "A novel form of the myeloid-specific zinc finger protein (MZF-2)." Genes to Cells. 2, (9). 581-591 (1997)

Murai, K.、Murakami, H. 和 Nagata, S.：“一种新形式的骨髓特异性锌指蛋白 (MZF-2)。”

Murakami, H.G-CSF,G-CSF receptor: Knockout mouse data book Edited by Kurokawa, K and Sasatsuki, T.Nakayama-Shoten, 194-195 (1997)

Murakami, H.G-CSF,G-CSF 受体：敲除小鼠数据书由 Kurokawa, K 和 Sasatsuki 编辑，T.Nakayama-Shoten，194-195 (1997)