Almeida E A, Ilić D, Han Q, Hauck C R, Jin F, Kawakatsu H, Schlaepfer D D, Damsky C H
Department of Stomatology and Department of Anatomy, Department of Medicine, University of California San Francisco, San Francisco, California 94143-0512, USA.
J Cell Biol. 2000 May 1;149(3):741-54. doi: 10.1083/jcb.149.3.741.
Most transformed cells have lost anchorage and serum dependence for growth and survival. Previously, we established that when serum is absent, fibronectin survival signals transduced by focal adhesion kinase (FAK), suppress p53-regulated apoptosis in primary fibroblasts and endothelial cells (Ilić et al. 1998. J. Cell Biol. 143:547-560). The present goals are to identify survival sequences in FAK and signaling molecules downstream of FAK required for anchorage-dependent survival of primary fibroblasts. We report that binding of the SH3 domain of p130Cas to proline-rich region 1 of FAK is required to support survival of fibroblasts on fibronectin when serum is withdrawn. The FAK-p130Cas complex activates c-Jun NH2-terminal kinase (JNK) via a Ras/Rac1/Pak1/MAPK kinase 4 (MKK4) pathway. Activated (phospho-) JNK colocalizes with FAK in focal adhesions of fibroblasts cultured on fibronectin, which supports their survival, but not in fibroblasts cultured on collagen, which does not. Cells often survive in the absence of extracellular matrix if serum factors are provided. In that case, we confirm work of others that survival signals are transduced by FAK, phosphatidylinositol 3'-kinase (PI3-kinase), and Akt/protein kinase B (PKB). However, when serum is absent, PI3-kinase and Akt/PKB are not involved in the fibronectin-FAK-JNK survival pathway documented herein. Thus, survival signals from extracellular matrix and serum are transduced by FAK via two distinct pathways.
大多数转化细胞已失去对生长和存活的锚定及血清依赖性。此前,我们证实,在无血清时,由粘着斑激酶(FAK)转导的纤连蛋白存活信号可抑制原代成纤维细胞和内皮细胞中p53调控的细胞凋亡(伊利奇等人,1998年。《细胞生物学杂志》143:547 - 560)。目前的目标是确定FAK中的存活序列以及原代成纤维细胞锚定依赖性存活所需的FAK下游信号分子。我们报告,当血清撤除时,p130Cas的SH3结构域与FAK富含脯氨酸区域1的结合是支持成纤维细胞在纤连蛋白上存活所必需的。FAK - p130Cas复合物通过Ras/Rac1/Pak1/丝裂原活化蛋白激酶激酶4(MKK4)途径激活c - Jun氨基末端激酶(JNK)。活化的(磷酸化的)JNK与FAK在纤连蛋白上培养的成纤维细胞的粘着斑中共定位,这支持它们的存活,但在胶原上培养的成纤维细胞中则不然,胶原上培养的细胞不能存活。如果提供血清因子,细胞在没有细胞外基质的情况下通常也能存活。在这种情况下,我们证实了其他人的研究结果,即存活信号由FAK、磷脂酰肌醇3'-激酶(PI3 -激酶)和Akt/蛋白激酶B(PKB)转导。然而,当无血清时,PI3 -激酶和Akt/PKB不参与本文所述的纤连蛋白 - FAK - JNK存活途径。因此,细胞外基质和血清的存活信号由FAK通过两条不同的途径转导。