The JNK1/JNK3 interactome - Contributions by the JNK3 unique N-terminus and JNK common docking site residues
The JNK1/JNK3 interactome - Contributions by the JNK3 unique N-terminus and JNK common docking site residues
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DOI：
10.1016/j.bbrc.2014.09.122
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发表时间：
2014-10-24
影响因子：
3.1
通讯作者：
Bogoyevitch, Marie A.
中科院分区：
文献类型：
--
作者：
Chen, Wei-Kai;Yeap, Yvonne Y. C.;Bogoyevitch, Marie A.
c-Jun N-末端激酶（JNK）在细胞对应激或生长因子刺激的反应中起重要作用。JNK 1 α 1同种型与主要神经元JNK 3 α 1同种型具有>90%的同一性，但JNK 3 α 1还包括独特的38个氨基酸的N-末端序列。为了解决这些JNK亚型具有介导不同生物学作用的不同结合伴侣的潜力这一悬而未决的问题，本文提出的工作改进了酵母双杂交方法，以鉴定和分类JNK 1 α 1和JNK 3 α 1的结合伴侣。具体而言，介导典型JNK结合结构域（JBD）依赖性相互作用的JNK 1 α 1共同对接（CD）结构域的定点诱变，独特的JNK 3 N末端结构域的截短，（即Δ N JNK 3 α 1），酵母双杂交系统中的相互作用评价将相互作用伴侣定义为JNK 1特异性相互作用物（ATF 7、FUS、KCNE 4、PIAS 1、SHANK 1、TKT），JNK 1 α 1和JNK 3 α 1共有的典型JBD依赖性相互作用物（AKAP 6、BMPR 2、EEF 1A 1、GFAP、GRIP 2、GTF 2F 1、HDAC 2、MAP 1B、MYO 9 B、PTPN 2、RABGAP 1、RUSC 2、SUMO 1、SYPL 1、TOPBP 1、ZNF 668）或JNK 3特异性伴侣（ATXN 1，NNAT，PTGDS）依赖于与JNK 3 N-末端延伸的相互作用。进一步探索了相互作用伙伴ATF 7、AKAP 6和ATXN 1作为这些不同类别的代表。在ATF 7中鉴定出两个潜在的JBD，其对于与JNK 1 α 1的相互作用是重要的，但另外，ATF 7与Delta N JNK 3 α 1之间的相互作用显示为JBD依赖性的，这表明JNK 3 α 1的N-末端阻止与某些蛋白质的相互作用。对于共享伴侣AKAP 6，通过序列分析预测的多个潜在JBD之一对于酵母筛选系统以及哺乳动物细胞中AKAP 6-JNK相互作用非常重要。最后，在哺乳动物细胞环境中，ATXN 1-JNK 3 α 1相互作用依赖于JNK 3 α 1 N-末端。因此，这些研究强调了潜在的JNK相互作用的合作伙伴与JBD依赖以及JBD独立的相互作用模式的多样性。(C)2014爱思唯尔公司All rights reserved.
The c-Jun N-terminal Kinases (JNKs) play important roles in cell responses to stress or growth factor stimulation. The JNK1 alpha 1 isoform shares >90% identity with a predominantly neuronal JNK3 alpha 1 isoform, but JNK3 alpha 1 also includes a distinctive 38 amino acid N-terminal sequence. To address the outstanding question of the potential for these JNK isoforms to have different binding partners that mediate different biological actions, the work presented here refined the yeast two-hybrid approach to identify and categorize binding partners for JNK1 alpha 1 and JNK3 alpha 1. Specifically, site-directed mutagenesis of the JNK1 alpha 1 common docking (CD) domain that mediates typical JNK-binding domain (JBD)-dependent interactions, truncation of the distinctive JNK3 N-terminal domain (i.e. Delta N JNK3 alpha 1), and interaction evaluation in the yeast two-hybrid system defined the interacting partners as either JNK1-specific interactors (ATF7, FUS, KCNE4, PIAS1, SHANK1, TKT), typical JBD-dependent interactors shared by JNK1 alpha 1 and JNK3 alpha 1 (AKAP6, BMPR2, EEF1A1, GFAP, GRIP2, GTF2F1, HDAC2, MAP1B, MYO9B, PTPN2, RABGAP1, RUSC2, SUMO1, SYPL1, TOPBP1, ZNF668), or JNK3-specific partners (ATXN1, NNAT, PTGDS) dependent on interaction with the JNK3 N-terminal extension. The interacting partners ATF7, AKAP6, and ATXN1 were explored further as representatives of these different classes. Two potential JBDs were identified in ATF7 as important for its interaction with JNK1 alpha 1, but additionally an interaction between ATF7 and Delta N JNK3 alpha 1 was shown to be JBD-dependent, suggesting that the JNK3 alpha 1 N-terminus prevents interaction with some proteins. For the shared partner AKAP6, one of the multiple potential JBDs predicted by sequence analysis was important for the AKAP6-JNK interaction in the yeast screening system as well as in mammalian cells. Finally, the ATXN1-JNK3 alpha 1 interaction was dependent on the JNK3 alpha 1 N-terminus in a mammalian cell context. These studies therefore highlight a diversity of potential JNK-interacting partners with both JBD-dependent as well as JBD-independent modes of interaction. (C) 2014 Elsevier Inc. All rights reserved.