c-Jun N-terminal kinase specifically phosphorylates p66ShcA at serine 36 in response to ultraviolet irradiation
c-Jun N-terminal kinase specifically phosphorylates p66ShcA at serine 36 in response to ultraviolet irradiation
复制标题
DOI：
10.1074/jbc.m106612200
复制
发表时间：
2001-12-21
影响因子：
4.8
通讯作者：
Maroney, AC
中科院分区：
文献类型：
--
作者：
Le, S;Connors, TJ;Maroney, AC
缺乏ShcA衔接蛋白p66异构体（p66(ShcA)）表达的Alice对氧化应激的敏感性较低，寿命延长。具体来说，p66（ShcA）丝氨酸36位点的磷酸化对于氧化损伤引起的细胞死亡反应至关重要。我们试图确定负责这种磷酸化的激酶。利用SH-SY5Y人神经母细胞瘤细胞模型，研究人员证实p66（ShcA）在紫外线照射下丝氨酸/苏氨酸残基磷酸化。c-Jun n末端激酶（JNKs）和p38丝裂原活化蛋白激酶都可以被紫外线照射激活，我们发现两者都能够在体外磷酸化p66（ShcA）的丝氨酸36。然而，在SH-SY5Y细胞中，用多谱系激酶抑制剂CEP-1347（可阻断紫外线诱导的JNK激活）而不是p38（磷脂酰肌醇3-激酶）或MEK1抑制剂处理细胞，可阻止p66（ShcA）磷酸化。与这一发现一致的是，转染激活的JNK1，而不是激酶死亡的JNK1，导致中国仓鼠卵巢细胞p66（ShcA）丝氨酸36磷酸化。综上所述，JNKs是SH-SY5Y细胞在紫外线照射下磷酸化p66（ShcA）丝氨酸36的激酶，通过干预JNK途径阻断p66（ShcA）磷酸化可以防止光诱导氧化应激引起的细胞损伤。
Alice lacking expression of the p66 isoform of the ShcA adaptor protein (p66(ShcA)) are less susceptible to oxidative stress and have an extended life span. Specifically, phosphorylation of p66(ShcA) at serine 36 is critical for the cell death response elicited by oxidative damage. We sought to identify the kinase(s) responsible for this phosphorylation. Utilizing the SH-SY5Y human neuroblastoma cell model, it is demonstrated that p66(ShcA) is phosphorylated on serine/threonine residues in response to UV irradiation. Both c-Jun N-terminal kinases (JNKs) and p38 mitogen-activated protein kinases are activated by UV irradiation, and we show that both are capable of phosphorylating serine 36 of p66(ShcA) in vitro. However, treatment of cells with a multiple lineage kinase inhibitor, CEP-1347, that blocks UV-induced JNK activation, but not p38, phosphatidylinositol 3-kinase, or MEK1 inhibitors, prevented p66(ShcA) phosphorylation in SH-SY5Y cells. Consistent with this finding, transfected activated JNK1, but not the kinase-dead JNK1, leads to phosphorylation of serine 36 of p66(ShcA) in Chinese hamster ovary cells. In conclusion, JNKs are the kinases that phosphorylate serine 36 of p66(ShcA) in response to UV irradiation in SH-SY5Y cells, and blocking p66(ShcA) phosphorylation by intervening in the JNK pathway may prevent cellular damage due to light-induced oxidative stress.