Diverse Sequence Determinants Control Human and Mouse Receptor Interacting Protein 3 (RIP3) and Mixed Lineage Kinase domain-Like (MLKL) Interaction in Necroptotic Signaling
Diverse Sequence Determinants Control Human and Mouse Receptor Interacting Protein 3 (RIP3) and Mixed Lineage Kinase domain-Like (MLKL) Interaction in Necroptotic Signaling
复制标题
多种序列决定因素控制坏死性凋亡信号传导中的人和小鼠受体相互作用蛋白 3 (RIP3) 和混合谱系激酶结构域样 (MLKL) 相互作用
DOI：
10.1074/jbc.m112.435545
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发表时间：
2013-06-07
影响因子：
4.8
通讯作者：
Han, Jiahuai
中科院分区：
文献类型：
--
作者：
Chen, Wanze;Zhou, Zhenru;Han, Jiahuai
受体相互作用蛋白3(RIP3)是一种在肿瘤坏死因子诱导的坏死性下垂过程中必不可少的蛋白激酶。人RIP3(HRIP3)中的Ser227蛋白(HRIP3)与肿瘤坏死因子刺激诱导的信号复合体中的人混合谱系激活区(MLKL)相互作用是必需的。RIP1和RIP3介导核小体聚集，导致淀粉样信号复合体的形成。我们发现，肿瘤坏死因子诱导小鼠RIP3(MRIP3)中Thr-231和Ser-232的磷酸化，并且这种磷酸化是mRIP3与mMLKL相互作用所必需的。MRIP3中的Ser-232对应于hRIP3中的Ser-227，而Thr-231在hRIP3中不保守。虽然RIP3-MLKL相互作用是人类和小鼠细胞发生坏死性下垂所必需的，但hRIP3不与mMLKL相互作用，mRIP3不能与hMLKL结合。RIP3-MLKL相互作用的物种特异性主要由hRIP3和mRIP3的磷酸化位点和磷酸化位点附近的侧翼序列的序列差异决定。尽管RIP3-MLKL相互作用在不同的生物体中同时出现了不同的结构和机制基础，但似乎RIP3-MLKL相互作用已被选为介导坏死性下垂信号的进化保守机制。此外，我们进一步揭示了RIP3与MLKL的相互作用阻止了大量异常的RIP3聚集，因此应该是形成死亡小体的淀粉样信号复合体的关键。我们还发现，RIP3和MLKL之间的相互作用是核小体移位到线粒体相关膜所必需的。我们的数据证明了RIP3-MLKL相互作用在功能性坏死体的形成中的重要性，并表明坏死体向线粒体相关膜的移位对于坏死性下垂信号的传递是必不可少的。
Receptor interacting protein 3 (RIP3) is a protein kinase essential for TNF-induced necroptosis. Phosphorylation on Ser227 in human RIP3 (hRIP3) is required for its interaction with human mixed lineage kinase domain-like (MLKL) in the necrosome, a signaling complex induced by TNF stimulation. RIP1 and RIP3 mediate necrosome aggregation leading to the formation of amyloid-like signaling complexes. We found that TNF induces Thr-231 and Ser-232 phosphorylation in mouse RIP3 (mRIP3) and this phosphorylation is required for mRIP3 to interact with mMLKL. Ser-232 in mRIP3 corresponds to Ser-227 in hRIP3, whereas Thr-231 is not conserved in hRIP3. Although the RIP3-MLKL interaction is required for necroptosis in both human and mouse cells, hRIP3 does not interact with mMLKL and mRIP3 cannot bind to hMLKL. The species specificity of the RIP3-MLKL interaction is primarily determined by the sequence differences in the phosphorylation sites and the flanking sequence around the phosphorylation sites in hRIP3 and mRIP3. It appears that the RIP3-MLKL interaction has been selected as an evolutionarily conserved mechanism in mediating necroptosis signaling despite that differing structural and mechanistic bases for this interaction emerged simultaneously in different organisms. In addition, we further revealed that the interaction of RIP3 with MLKL prevented massive abnormal RIP3 aggregation, and therefore should be crucial for formation of the amyloid signaling complex of necrosomes. We also found that the interaction between RIP3 and MLKL is required for the translocation of necrosomes to mitochondria-associated membranes. Our data demonstrate the importance of the RIP3-MLKL interaction in the formation of functional necrosomes and suggest that translocation of necrosomes to mitochondria-associated membranes is essential for necroptosis signaling.