Zamorano J, Keegan A D
Department of Immunology, Jerome Holland Laboratories, American Red Cross, Rockville, MD 20855, USA.
J Immunol. 1998 Jul 15;161(2):859-67.
IL-4 is a cytokine with important antiapoptotic activity. We have analyzed the role that tyrosine-containing domains within the cytoplasmic tail of IL-4R alpha play in IL-4-mediated protection from apoptosis. 32D cells expressing a wt huIL-4R alpha or one truncated at aa 557 were protected by huIL-4 from apoptosis while cells expressing a receptor truncated at aa 657 were not, suggesting that the carboxyl-terminal domain signals protection from apoptosis. However, changing Y713 within this region to phenylalanine had no effect. To analyze the contribution of tyrosine-containing domains independently, we transplanted regions of the huIL-4R alpha to a truncated form of the huIL-2R beta that could not signal protection from apoptosis. Transplantation of the huIL-4R alpha domains containing Y497 or Y713 partially prevented cell death and together signaled protection from apoptosis in response to IL-2 as well as the wt IL-2R beta. Mutation of Y497 and Y713 to phenylalanine inhibited protection. In contrast, transplantation of the domain containing the potential STAT6-docking tyrosines alone had no effect, yet it inhibited the protection mediated by the other domains. Although IL-4R alpha signals Shc and SH2-containing inositol phosphatase (SHIP) phosphorylation, we could not establish an association between their activation and protection from apoptosis. Taken together, this study suggests that the domains of the huIL-4R alpha containing Y497 and Y713 positively regulate protection from apoptosis while the domain containing the STAT6 docking sites suppresses this protection, and that additional signaling molecules other than insulin receptor substrate-1 (IRS1), Shc, or SHIP may be involved in antiapoptotic signaling.
白细胞介素-4(IL-4)是一种具有重要抗凋亡活性的细胞因子。我们分析了IL-4Rα细胞质尾部含酪氨酸结构域在IL-4介导的抗凋亡保护中所起的作用。表达野生型人IL-4Rα或在第557位氨基酸处截短的32D细胞受到人IL-4的抗凋亡保护,而表达在第657位氨基酸处截短的受体的细胞则没有,这表明羧基末端结构域发出抗凋亡保护信号。然而,将该区域内的Y713突变为苯丙氨酸没有影响。为了独立分析含酪氨酸结构域的作用,我们将人IL-4Rα的区域移植到不能发出抗凋亡保护信号的人IL-2Rβ截短形式上。含有Y497或Y713的人IL-4Rα结构域的移植部分地阻止了细胞死亡,并共同发出了对IL-2以及野生型IL-2Rβ抗凋亡保护的信号。将Y497和Y713突变为苯丙氨酸会抑制保护作用。相反,单独移植含有潜在STAT6对接酪氨酸的结构域没有作用,但它会抑制其他结构域介导的保护作用。尽管IL-4Rα可使含Shc和含SH2的肌醇磷酸酶(SHIP)磷酸化,但我们无法确定它们的激活与抗凋亡保护之间的关联。综上所述,本研究表明,含有Y497和Y7由3个数字组成,分别是713的人IL-4Rα结构域正向调节抗凋亡保护,而含有STAT6对接位点的结构域则抑制这种保护,并且除胰岛素受体底物-1(IRS1)、Shc或SHIP之外,可能还有其他信号分子参与抗凋亡信号传导。