Tyrosine residues of the granulocyte colony-stimulating factor receptor transmit proliferation and differentiation signals in murine bone marrow cells
Tyrosine residues of the granulocyte colony-stimulating factor receptor transmit proliferation and differentiation signals in murine bone marrow cells
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DOI：
10.1182/blood.v99.3.879
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发表时间：
2002-02-01
期刊：
影响因子：
20.3
通讯作者：
Layton, JE
中科院分区：
文献类型：
--
作者：
Akbarzadeh, S;Ward, AC;Layton, JE
粒细胞集落刺激因子 (G-CSF) 是粒细胞生成的主要调节因子，通过与其在中性粒细胞上的特异性受体 (G-CSF-R) 结合发挥作用。先前对 4 G-CSF-R 细胞质酪氨酸残基信号传导的研究使用的模型细胞系可能具有特殊的非生理反应。本研究旨在鉴定原代骨髓细胞中受体酪氨酸残基传递的特定信号。为了绕过内源性G-CSF-R的存在，使用含有表皮生长因子受体胞外域的嵌合受体代替G-CSF-R的整个胞外域。构建了一系列单独或共同含有酪氨酸苯丙氨酸突变的嵌合受体，并在 G-CSF 缺陷小鼠的原代骨髓细胞中表达。测量表皮生长因子刺激的受体表达骨髓细胞的增殖和分化反应。增加 50% 的有效浓度来刺激受体 Y 无效突变体表明细胞增殖需要来自酪氨酸残基的特定信号，特别是在低浓度的刺激下。突变受体的反应受损表明 G-CSF-R Y-764 参与细胞增殖，Y-729 参与粒细胞分化信号传导。此外，突变受体之间对配体刺激的不同敏感性表明G-CSF-R Y-744和可能的Y-729在细胞增殖中具有抑制作用。 STAT 激活不受酪氨酸突变的影响，而 ERK 激活似乎至少部分依赖于 Y-764。这些观察结果表明原代细胞中 G-CSF-R 酪氨酸残基具有新的作用，这是以前在细胞系研究中未观察到的。 (C) 2002 年，美国血液学会。
Granulocyte colony-stimulating factor (G-CSF) is the major regulator of granulopoiesis and acts through binding to its specific receptor (G-CSF-R) on neutrophilic granulocytes. Previous studies of signaling from the 4 G-CSF-R cytoplasmic tyrosine residues used model cell lines that may have idiosyncratic, nonphysiological responses. This study aimed to identify specific signals transmitted by the receptor tyrosine residues in primary myeloid cells. To bypass the presence of endogenous G-CSF-R, a chimeric receptor containing the extracellular domain of the epidermal growth factor receptor in place of the entire extracellular domain of the G-CSF-R was used. A series of chimeric receptors containing tyrosine mutations to phenylalanine, either individually or collectively, was constructed and expressed in primary bone marrow cells from G-CSF-deficient mice. Proliferation and differentiation responses of receptor-expressing bone marrow cells stimulated by epidermal growth factor were measured. An increased 50% effective concentration to stimulus of the receptor Y-null mutant indicated that specific signals from tyrosine residues were required for cell proliferation, particularly at low concentrations of stimulus. Impaired responses by mutant receptors implicated G-CSF-R Y-764 in cell proliferation and Y-729 in granulocyte differentiation signaling. In addition, different sensitivities to ligand stimulation between mutant receptors indicated that G-CSF-R Y-744 and possibly Y-729 have an inhibitory role in cell proliferation. STAT activation was not affected by tyrosine mutations, whereas ERK activation appeared to depend, at least in part, on Y-764. These observations have suggested novel roles for the G-CSF-R tyrosine residues in primary cells that were not observed previously in studies in cell lines. (C) 2002 by The American Society of Hematology.