Protein kinase PKN1 associates with TRAF2 and is involved in TRAF2-NF-κB signaling pathway
Protein kinase PKN1 associates with TRAF2 and is involved in TRAF2-NF-κB signaling pathway
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DOI：
10.1016/j.bbrc.2003.12.148
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发表时间：
2004-02-13
影响因子：
3.1
通讯作者：
Ono, Y
中科院分区：
文献类型：
--
作者：
Gotoh, Y;Oishi, K;Ono, Y
PKN 1是一种脂肪酸和Rho激活的丝氨酸/苏氨酸蛋白激酶，其催化结构域与蛋白激酶C（PKC）家族高度同源。在PKN 1结合蛋白的酵母双杂交筛选中，我们鉴定了肿瘤坏死因子α（TNF α）受体相关因子2（TRAF 2）。TRAF 2是TNF受体超家族的主要介导因子之一，其将TNF信号转导至各种功能靶点，包括激活NF-κ B、JNK和凋亡。FLAG标记的PKN 1与来自HEK 293细胞裂解物的内源性TRAF 2共免疫沉淀，并且使用TRAF 2的缺失突变体的体外结合测定显示PKN 1直接结合TRAF 2的TRAF结构域。PKN 1在氨基酸残基580-584（PIQES）处具有TRAF 2结合共有序列PXQX（S/T），并且P580 AQ 582 A突变体不与TRAF 2共免疫沉淀。此外，通过RNA干扰（RNAi）降低PKN 1的表达下调了HEK 293 T细胞中TRAF 2诱导的NF-κ B活化。这些结果表明PKN 1参与TRAF 2-NF-kappaB信号通路。(C)2003年爱思唯尔公司All rights reserved.
PKN1 is a fatty acid and Rho-activated serine/threonine protein kinase whose catalytic domain is highly homologous to protein kinase C (PKC) family. In yeast two-hybrid screening for PKN1 binding proteins, we identified tumor necrosis factor alpha (TNFalpha) receptor-associated factor 2 (TRAF2). TRAF2 is one of the major mediators of TNF receptor superfamily transducing TNF signal to various functional targets, including activation of NF-kappaB, JNK, and apoptosis. FLAG-tagged PKN1 was co-immunoprecipitated with endogenous TRAF2 from HEK293 cell lysate, and in vitro binding assay using the deletion mutants of TRAF2 showed that PKN1 directly binds to the TRAF domain of TRAF2. PKN1 has the TRAF2-binding consensus sequences PXQX (S/T) at amino acid residues 580-584 (PIQES), and P580AQ582A mutant was not co-immunoprecipitated with TRAF2. Furthermore, the reduced expression of PKN1 by RNA interference (RNAi) down-regulated TRAF2-induced NF-kappaB activation in HEK293T cells. These results suggest that PKN1 is involved in TRAF2-NF-kappaB signaling pathway. (C) 2003 Elsevier Inc. All rights reserved.