Signal transduction mechanisms of neutrophil differentiation through G-CSF receptor.

通过G-CSF受体的中性粒细胞分化的信号转导机制。

• 批准号: 14580700
• 负责人: MURAKAMI Hiroshi
• 金额: $ 1.98万
• 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: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580700/
• 关键词: signal transduction; colony stimulating factor; receptor; proliferation; differentiation; neutrophil; cytokine; granulocyte

G-CSF stimulation leads to the myeloid precursor cells to proliferate for a few days, then they stop proliferating, followed by differentiation to the mature neutrophils. The tyrosine residues in membrane-distal region of the G-CSF receptor as well as the activation of STAT3 was involved in the growth arrest during the neutrophil differentiation. However, involvement of other residues of the receptor and other signaling molecules in the growth arrest remains to be determine. We introduced mutations in the cytoplasmic region of the receptor and transfected them into granulocyte precursor cells which did not express endogenous G-CSF receptor. A cell line was isolated that proliferated continuously in the presence of G-CSF, that is, that did not show growth-arrest in the medium containing G-CSF. The mutated G-CSF receptor genes were isolated by PCR from the chromosomal DNA of the obtained cell line. Each mutant gene was re-introduced into the granulocyte precursor cells and G-CSF-response … More s of the obtained stable transformants were examined. A cell line which proliferated continuously in the medium with G-CSF, carved G-CSF receptor with its 59 amino acid deletion in its C-terminus, suggesting this C-terminus domain is responsible for transducing the growth arrest signals. A series of truncated receptors were constructed and cell lines expressing these truncated receptors were established. G-CSF-responses of the cell lines harboring these truncated receptors revealed that C-terminal 25 amino acid residues of the G-CSF receptor was necessary for the transducing growth arrest signals. Cells expressing the truncated receptors also showed the defects in the G-CSF-dependent induction of neutrophilic morphological changes with lobulated nucleus. The G-CSF dependent activation of STATS was prolonged in the cells expressing the truncated receptor. Therefore, the C-terminal 25 amino acid residues were responsible for the sustained activation of STAT5, resulting in the defects in the G-CSF dependent growth arrest and morphological changes. Less

G-CSF刺激导致髓样前体细胞增殖数天，然后它们停止增殖，随后分化为成熟的中性粒细胞。G-CSF受体膜远端区的酪氨酸残基以及STAT 3的激活参与了中性粒细胞分化过程中的生长停滞。然而，参与生长停滞的受体和其他信号分子的其他残基仍有待确定。我们在受体的胞质区域引入突变，并将其转染到不表达内源性G-CSF受体的粒细胞前体细胞中。分离出在G-CSF存在下连续增殖的细胞系，即，在含有G-CSF的培养基中不显示生长停滞。通过PCR从获得的细胞系的染色体DNA中分离突变的G-CSF受体基因。将每个突变基因重新导入粒细胞前体细胞， ...更多信息 对获得的稳定转化体进行检测。在含有G-CSF的培养基中连续增殖的细胞系，在其C-末端切割G-CSF受体，其59个氨基酸缺失，表明该C-末端结构域负责转导生长停滞信号。构建了一系列截短受体，并建立了表达这些截短受体的细胞系。携带这些截短的受体的细胞系的G-CSF-应答揭示G-CSF受体的C-末端25个氨基酸残基对于转导生长停滞信号是必需的。表达截短受体的细胞也表现出G-CSF依赖性诱导的具有分叶核的嗜酸性形态学变化的缺陷。在表达截短受体的细胞中，G-CSF依赖的STATS活化延长。因此，C端25个氨基酸残基负责STAT 5的持续激活，导致G-CSF依赖的生长停滞和形态学变化的缺陷。少

项目成果

村上 宏: "生物工学会誌 第82巻 第4号"日本生物工学会(印刷中). (2004)

村上浩：《生物工程学会杂志》第 82 卷第 4 期》日本生物工程学会（出版中）（2004 年）。

Hiroshi Murakami: "Neurotrophil differentiation with morphological changes by G-CSF stimulation."Seibutsu-Kogaku Kaishi (Japanese). Vol.4(in press). (2004)

Hiroshi Murakami：“G-CSF 刺激引起的神经粒细胞分化和形态变化。”Seibutsu-Kogaku Kaishi（日语）。

Omura, T., et al.: "Acceleration of granulocyte colony-stimulating factor-induced neutrophilic nuclear lobulation by overexpression of Lyn tyrosine kinase"European J. Biochem. 269. 381-389 (2002)

Omura，T.等人：“通过Lyn酪氨酸激酶的过度表达加速粒细胞集落刺激因子诱导的中性粒细胞核分叶”European J.Biochem。