Senis Y, Zirngibl R, McVeigh J, Haman A, Hoang T, Greer P A
Department of Pathology, Cancer Research Laboratories, Queen's University, Kingston, Ontario K7L 3N6.
Mol Cell Biol. 1999 Nov;19(11):7436-46. doi: 10.1128/MCB.19.11.7436.
The fps/fes proto-oncogene encodes a cytoplasmic protein-tyrosine kinase that is functionally implicated in the survival and terminal differentiation of myeloid progenitors and in signaling from several members of the cytokine receptor superfamily. To gain further insight into the physiological function of fps/fes, we targeted the mouse locus with a kinase-inactivating missense mutation. Mutant Fps/Fes protein was expressed at normal levels in these mice, but it lacked detectable kinase activity. Homozygous mutant animals were viable and fertile, and they showed no obvious defects. Flow cytometry analysis of bone marrow showed no statistically significant differences in the levels of myeloid, erythroid, or B-cell precursors. Subtle abnormalities observed in mutant mice included slightly elevated total leukocyte counts and splenomegaly. In bone marrow hematopoietic progenitor cell colony-forming assays, mutant mice gave slightly elevated numbers and variable sizes of CFU-granulocyte macrophage in response to interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF). Tyrosine phosphorylation of Stat3 and Stat5A in bone marrow-derived macrophages was dramatically reduced in response to GM-CSF but not to IL-3 or IL-6. This suggests a distinct nonredundant role for Fps/Fes in signaling from the GM-CSF receptor that does not extend to the closely related IL-3 receptor. Lipopolysaccharide-induced Erk1/2 activation was also reduced in mutant macrophages. These subtle molecular phenotypes suggest a possible nonredundant role for Fps/Fes in myelopoiesis and immune responses.
fps/fes原癌基因编码一种细胞质蛋白酪氨酸激酶,其在髓系祖细胞的存活和终末分化以及细胞因子受体超家族的几个成员的信号传导中具有功能作用。为了进一步深入了解fps/fes的生理功能,我们用激酶失活的错义突变靶向小鼠基因座。突变的Fps/Fes蛋白在这些小鼠中以正常水平表达,但缺乏可检测到的激酶活性。纯合突变动物存活且可育,未表现出明显缺陷。骨髓的流式细胞术分析显示,髓系、红系或B细胞前体水平无统计学显著差异。在突变小鼠中观察到的细微异常包括总白细胞计数略有升高和脾肿大。在骨髓造血祖细胞集落形成试验中,突变小鼠对白细胞介素-3(IL-3)和粒细胞-巨噬细胞集落刺激因子(GM-CSF)的反应中,CFU-粒细胞巨噬细胞的数量略有增加且大小不一。骨髓来源的巨噬细胞中,Stat3和Stat5A的酪氨酸磷酸化在对GM-CSF的反应中显著降低,但对IL-3或IL-6的反应中未降低。这表明Fps/Fes在GM-CSF受体信号传导中具有独特的非冗余作用,该作用并不延伸至密切相关的IL-3受体。突变巨噬细胞中脂多糖诱导的Erk1/2激活也降低。这些细微的分子表型表明Fps/Fes在骨髓生成和免疫反应中可能具有非冗余作用。