Sun Yue, Cheng Zhijie, Ma Lan, Pei Gang
Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, People's Republic of China.
J Biol Chem. 2002 Dec 20;277(51):49212-9. doi: 10.1074/jbc.M207294200. Epub 2002 Oct 4.
Chemotaxis mediated by chemokine receptors such as CXCR4 plays a key role in lymphocyte homing and hematopoiesis as well as in breast cancer metastasis. We have demonstrated previously that beta-arrestin2 functions to attenuate CXCR4-mediated G protein activation and to enhance CXCR4 internalization. Here we show further that the expression of beta-arrestin2 in both HeLa and human embryonic kidney 293 cells significantly enhances the chemotactic efficacy of stromal cell-derived factor 1alpha, the specific agonist of CXCR4, whereas the suppression of beta-arrestin2 endogenous expression by antisense or RNA-mediated interference technology considerably attenuates stromal cell-derived factor 1alpha-induced cell migration. Expression of beta-arrestin2 also augmented chemokine receptor CCR5-mediated but not epidermal growth factor receptor-mediated chemotaxis, indicating the specific effect of beta-arrestin2. Further analysis reveals that expression of beta-arrestin2 strengthened CXCR4-mediated activation of both p38 MAPK and ERK, and the suppression of beta-arrestin2 expression blocked the activation of two kinases. Interestingly, inhibition of p38 MAPK activation (but not ERK activation) by its inhibitors or by expression of a dominant-negative mutant of p38 MAPK effectively blocked the chemotactic effect of beta-arrestin2. Expression of a dominant-negative mutant of ASK1 also exerted the similar blocking effect. The results of our study suggest that beta-arrestin2 can function not only as a regulator of CXCR4 signaling but also as a mediator of stromal cell-derived factor 1alpha-induced chemotaxis and that this activity probably occurs via the ASK1/p38 MAPK pathway.
由趋化因子受体(如CXCR4)介导的趋化作用在淋巴细胞归巢、造血作用以及乳腺癌转移过程中发挥着关键作用。我们之前已经证明,β -抑制蛋白2能够减弱CXCR4介导的G蛋白激活,并增强CXCR4的内化作用。在此我们进一步表明,在人宫颈癌HeLa细胞和人胚肾293细胞中,β -抑制蛋白2的表达均显著增强了基质细胞衍生因子1α(CXCR4的特异性激动剂)的趋化功效,而通过反义或RNA介导的干扰技术抑制β -抑制蛋白2的内源性表达,则会显著减弱基质细胞衍生因子1α诱导的细胞迁移。β -抑制蛋白2的表达还增强了趋化因子受体CCR5介导的趋化作用,但对表皮生长因子受体介导的趋化作用没有影响,这表明β -抑制蛋白2具有特异性作用。进一步分析显示,β -抑制蛋白2的表达增强了CXCR4介导的p38丝裂原活化蛋白激酶(p38 MAPK)和细胞外信号调节激酶(ERK)的激活,而抑制β -抑制蛋白2的表达则会阻断这两种激酶的激活。有趣的是,通过其抑制剂或表达p38 MAPK的显性负性突变体抑制p38 MAPK的激活(而非ERK的激活),能够有效阻断β -抑制蛋白2的趋化作用。表达ASK1的显性负性突变体也产生了类似的阻断作用。我们的研究结果表明,β -抑制蛋白2不仅可以作为CXCR4信号的调节剂,还可以作为基质细胞衍生因子1α诱导趋化作用的介质,并且这种活性可能是通过ASK1/p38 MAPK途径发生的。