Yamada Satomi, Marutsuka Masaki, Inoue Miyabi, Zhang Jidong, Abe Shin-Ichi, Ishibashi Ken-Ichi, Yamaguchi Naoto, Eto Ko
Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan.
Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University 1-8-1 Inohana, Chuo-ku, Chiba 260-8675, Japan.
Int J Biochem Mol Biol. 2014 May 15;5(1):21-9. eCollection 2014.
We have shown that the receptor tyrosine kinase ErbB4 signals neuregulin1-stimulated proliferation of human cells. Some isoforms of ErbB4 are cleaved to release the soluble intracellular domain p80; however, the function of p80 in cell proliferation remained unclear. Here we propose the possibility for p80 as a negative feedback modulator of ErbB4-mediated cell proliferation. Cells exposed to lower doses of neuregulin1 displayed a stimulated proliferation and contained ErbB4 but barely p80. By contrast, cells exposed to its higher doses displayed a suppressed proliferation and contained p80 but barely ErbB4. Analyses with cells overexpressing the p80 wild type and mutants indicated that nuclear p80 inhibits cell proliferation independently of the tyrosine kinase activity. A screen for a novel protein that interacts with p80 identified α-enolase, which is reported as a transcriptional inhibitor for the proliferation-associated c-myc gene. The c-myc mRNA expression was induced by lower doses of neuregulin1 but was suppressed by its higher doses. Subcellular fractionation demonstrated the localization of not only p80 and α-enolase but also the decrease of the functional c-myc amount in the nuclei of cells exposed to higher doses of neuregulin1. These results suggested that p80, which is generated from ErbB4 and translocates to the nuclei, interacts with α-enolase and inhibits neuregulin1-dependent ErbB4-mediated cell proliferation by impairing the c-myc gene transcription.
我们已经证明,受体酪氨酸激酶ErbB4可传导神经调节蛋白1刺激的人类细胞增殖信号。ErbB4的一些亚型会被切割,释放出可溶性细胞内结构域p80;然而,p80在细胞增殖中的功能仍不清楚。在此,我们提出p80作为ErbB4介导的细胞增殖的负反馈调节因子的可能性。暴露于较低剂量神经调节蛋白1的细胞表现出增殖受刺激,含有ErbB4,但几乎不含p80。相比之下,暴露于较高剂量神经调节蛋白1的细胞表现出增殖受抑制,含有p80,但几乎不含ErbB4。对过表达p80野生型和突变体的细胞进行分析表明,细胞核中的p80独立于酪氨酸激酶活性抑制细胞增殖。对一种与p80相互作用的新型蛋白质的筛选鉴定出α-烯醇化酶,据报道它是增殖相关c-myc基因的转录抑制剂。较低剂量的神经调节蛋白1可诱导c-myc mRNA表达,但其较高剂量则会抑制该表达。亚细胞分级分离表明,在暴露于较高剂量神经调节蛋白1的细胞的细胞核中,不仅有p80和α-烯醇化酶的定位,而且功能性c-myc的量也减少。这些结果表明,由ErbB4产生并转运至细胞核的p80与α-烯醇化酶相互作用,并通过损害c-myc基因转录来抑制神经调节蛋白1依赖的ErbB4介导的细胞增殖。