The tandem EH domains of End3 cooperate to interact with dual XPF motifs of Sla1 for the connection of early and late stages in fungal endocytosis
The tandem EH domains of End3 cooperate to interact with dual XPF motifs of Sla1 for the connection of early and late stages in fungal endocytosis
复制标题
DOI：
10.1016/j.bbrc.2023.04.075
复制
发表时间：
2023-04-28
影响因子：
3.1
通讯作者：
Wang，Dongli
中科院分区：
文献类型：
--
作者：
Zhang，Xiaokang;Wang，Chao;Wang，Dongli
网格蛋白介导的内吞作用（CME）是真核细胞生理过程中必不可少的。在真菌中，Pan 1/End 3/Sla 1复合物控制CME早期和晚期之间的过渡。End 3是利用其N端与Sla 1的C端相互作用的蛋白质，但其具体作用机制尚不清楚。稻瘟病菌（Magnaporthe bacteria）是水稻的病原真菌，引起稻瘟病，严重威胁着世界水稻生产。本文详细报道了M. martensa的End 3和Sla 1之间的相互作用机制，即MoEnd 3和MoSla 1。MoEnd 3的两个EH结构域（MoEnd 3-EH 1和MoEnd 3-EH 2）在进化和钙结合方面都是不同的，但对于彼此的构象稳定性是不可或缺的，串联排列的EH结构域的未报道的效果。MoEnd 3-EH 1和MoEnd 3-EH 2以保守模式与含有NPF基序的肽MoSla 11145 − 1155相互作用，MoEnd 3-EH（含有EH 1和EH 2结构域）以更高的亲和力与MoSla 11145 − 1155结合，支持EH结构域的协同效应。此外，MoEnd 3-EH还识别具有新的MPF基序的肽MoSla 1971 - 981，而酵母的Sla 1含有具有EH结构域相互作用能力的DPF基序。总的来说，我们的研究表明，End 3的两个EH结构域协同作用，与Sla 1的双XPF基序相互作用，这符合二价受体-二价配体模型，以提高亲和力和特异性。
Clathrin-mediated endocytosis (CME) is imperative for physiological processes in eukaryotic cells. In fungi, the Pan1/End3/Sla1 complex controls the transition between early and late stages of CME. Although it is acknowledged that End3 uses its N-terminal to interact with the C-terminal of Sla1, detailed mechanism remains obscure.Magnaporthe oryzae, the pathogenic fungus of rice, cause blast disease that threatens rice production worldwide. Here we report the detailed interaction mechanism between End3 and Sla1 ofM. oryzae, i.e. MoEnd3 and MoSla1. The two EH domains of MoEnd3 (MoEnd3-EH1 and MoEnd3-EH2) is different both in evolution and calcium binding, but are indispensable for conformational stability of each other, an unreported effect of tandem-arranged EH domains. MoEnd3-EH1 and MoEnd3-EH2 interact with peptide MoSla11145−1155containing a NPF motif with a conserved mode, and MoEnd3-EHs (containing both EH1 and EH2 domains) binds MoSla11145−1155with a higher affinity, supporting the synergetic effect of EH domains. In addition, MoEnd3-EHs also recognize peptide MoSla1971−981with a new MPF motif that has not been reported before, while Sla1 of yeast contains a DPF motif that bears EH domain interaction ability. Collectively, our research shows that the two EH domains of End3 synergize to interact with dual XPF motifs of Sla1, which conforms to a bivalent receptor-bivalent ligand model to improve both affinity and specificity.