Lin T H, Rosales C, Mondal K, Bolen J B, Haskill S, Juliano R L
Department of Pharmacology, School of Medicine, University of North Carolina, Chapel Hill 27599, USA.
J Biol Chem. 1995 Jul 7;270(27):16189-97. doi: 10.1074/jbc.270.27.16189.
Activation of cytoplasmic tyrosine kinases is an important aspect of signal transduction mediated by integrins. In the human monocytic cell line THP-1, either integrin-dependent cell adhesion to fibronectin or ligation of beta 1 integrins with antibodies causes a rapid and intense tyrosine phosphorylation of two sets of proteins of about 65-75 and 120-125 kDa. In addition, integrin ligation leads to nuclear translocation of the p50 and p65 subunits of the NF-kappa B transcription factor, to activation of a reporter gene driven by a promoter containing NF-kappa B sites, and to increased levels of mRNAs for immediate-early genes, including the cytokine interleukin (IL)-1 beta. The tyrosine kinase inhibitors genistein and herbimycin A block both integrin-mediated tyrosine phosphorylation and increases in IL-1 beta message levels, indicating a causal relationship between the two events. The components tyrosine phosphorylated subsequent to cell adhesion include paxillin, pp125FAK, and the SH2 domain containing tyrosine kinase Syk. In contrast, integrin ligation with antibodies induces tyrosine phosphorylation of Syk but not of FAK or paxillin. In adhering cells, pre-treatment with cytochalasin D suppresses tyrosine phosphorylation of FAK and paxillin but not of Syk, while IL-1 beta message induction is unaffected. These observations indicate that the Syk tyrosine kinase may be an important component of an integrin signaling pathway in monocytic cells, leading to activation of NF-kappa B and to increased levels of cytokine messages.
细胞质酪氨酸激酶的激活是整合素介导的信号转导的一个重要方面。在人单核细胞系THP-1中,整合素依赖的细胞与纤连蛋白的黏附或用抗体连接β1整合素都会导致两组分子量约为65 - 75 kDa和120 - 125 kDa的蛋白质快速且强烈的酪氨酸磷酸化。此外,整合素连接导致NF-κB转录因子的p50和p65亚基向细胞核转位,激活由含有NF-κB位点的启动子驱动的报告基因,并导致包括细胞因子白细胞介素(IL)-1β在内的立即早期基因的mRNA水平升高。酪氨酸激酶抑制剂染料木黄酮和除莠霉素A可阻断整合素介导的酪氨酸磷酸化以及IL-1β信使水平的升高,表明这两个事件之间存在因果关系。细胞黏附后酪氨酸磷酸化的成分包括桩蛋白、pp125FAK和含SH2结构域的酪氨酸激酶Syk。相反,用抗体连接整合素可诱导Syk的酪氨酸磷酸化,但不诱导FAK或桩蛋白的酪氨酸磷酸化。在黏附细胞中,用细胞松弛素D预处理可抑制FAK和桩蛋白的酪氨酸磷酸化,但不抑制Syk的酪氨酸磷酸化,而IL-1β信使诱导不受影响。这些观察结果表明,Syk酪氨酸激酶可能是单核细胞中整合素信号通路的一个重要组成部分,导致NF-κB激活和细胞因子信使水平升高。