Bombesin, lysophosphatidic acid, and epidermal growth factor rapidly stimulate focal adhesion kinase phosphorylation at Ser-910 - Requirement for ERK activation
Bombesin, lysophosphatidic acid, and epidermal growth factor rapidly stimulate focal adhesion kinase phosphorylation at Ser-910 - Requirement for ERK activation
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DOI：
10.1074/jbc.m210876200
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发表时间：
2003-06-20
影响因子：
4.8
通讯作者：
Rozengurt, E
中科院分区：
文献类型：
--
作者：
Hunger-Glaser, I;Salazar, EP;Rozengurt, E
在多个信号分子刺激的细胞中，粘着斑激酶(FAK)的酪氨酸磷酸化迅速增加，但对FAK在丝氨酸残基的磷酸化调控几乎一无所知。蛙皮素对瑞士3T3细胞的刺激促进了Ser-910处FAK的磷酸化显著增加(类似于13倍)，识别该残基的磷酸化状态的位点特异性抗体揭示了这一点。溶血磷脂酸和表皮生长因子(EGF)也可刺激Ser-910上FAK的磷酸化。佛波醇-12，13-二丁酸酯(PDB)对蛋白激酶C亚型的直接激活也促进了FAK在Ser-910的显著磷酸化。用蛋白激酶C抑制剂GF I或RO31-8220处理或长期暴露于PDB可阻止蛙皮素或PDB诱导的Ser-910 FAK磷酸化增加，但不能阻止EGF的作用。ERK抑制剂U0126和PD98059的处理阻止了FAK在Ser-910处的磷酸化，以响应所有测试的刺激。此外，在体外，激活的ERK2与FAK免疫复合物孵育导致FAK在Ser-910处的磷酸化。我们的结果首次证明，蛙皮素、溶血磷脂酸、PDB或EGF刺激可通过ERK依赖的途径在瑞士3T3细胞中诱导内源性FAK在Ser-910处的磷酸化。
A rapid increase in the tyrosine phosphorylation of focal adhesion kinase (FAK) has been extensively documented in cells stimulated by multiple signaling molecules, but virtually nothing is known about the regulation of FAK phosphorylation at serine residues. Stimulation of Swiss 3T3 cells with bombesin promoted a striking increase (similar to13-fold) in the phosphorylation of FAK at Ser-910, as revealed by site-specific antibodies that recognized the phosphorylated state of this residue. Lysophosphatidic acid and epidermal growth factor (EGF) also stimulated FAK phosphorylation at Ser-910. Direct activation of protein kinase C isoforms with phorbol-12,13-dibutyrate (PDB) also promoted striking phosphorylation of FAK at Ser-910. Treatment with the protein kinase C inhibitor GF I or Ro 31-8220 or chronic exposure to PDB prevented the increase in FAK phosphorylation at Ser-910 induced by bombesin or PDB but not by EGF. Treatment with the ERK inhibitors U0126 and PD98059 prevented FAK phosphorylation at Ser-910 in response to all of the stimuli tested. Furthermore, incubation of activated ERK2 with FAK immunocomplexes leads to FAK phosphorylation at Ser-910 in vitro. Our results demonstrate, for the first time, that stimulation with bombesin, lysophosphatidic acid, PDB, or EGF induces phosphorylation of endogenous FAK at Ser-910 via an ERK-dependent pathway in Swiss 3T3 cells.