Genetically incorporated crosslinkers reveal NleE attenuates host autophagy dependent on PSMD10.
Genetically incorporated crosslinkers reveal NleE attenuates host autophagy dependent on PSMD10.
复制标题
基因整合的交联剂揭示 NleE 减弱依赖于 PSMD10 的宿主自噬
作者：
Li J;Guo S;Chai F;Sun Q;Li P;Gao L;Dai L;Ouyang X;Zhou Z;Zhou L;Cheng W;Qi S;Lu K;Ren H
自噬是对抗感染的关键先天免疫反应。一些毒力效应子会破坏宿主的自噬机制以逃避自噬的监视。病原体与宿主自噬相互作用的机制仍不清楚。然而，由于这些相互作用的弱性、动态性和瞬时性，传统策略通常难以识别与效应子相互作用的宿主蛋白。在这里，我们发现肠致病性大肠杆菌 (EPEC) 依赖效应子 NleE 调节宿主细胞中自噬体的形成。通过采用基因整合的交联剂，26S 蛋白酶体调节亚基 10 (PSMD10) 被鉴定为活细胞中 NleE 的直接相互作用伙伴。成对化学交联揭示 NleE 与 PSMD10 的 N 末端相互作用。我们证明 PSMD10 同二聚化对于其与 ATG7 相互作用和促进自噬是必需的，但对于 PSMD10 与 ATG12 相互作用不是必需的。因此，单体状态下的 NleE 介导的 PSMD10 会减弱宿主自噬体的形成。我们的研究揭示了 EPEC 减弱宿主自噬活性的机制。
Autophagy acts as a pivotal innate immune response against infection. Some virulence effectors subvert the host autophagic machinery to escape the surveillance of autophagy. The mechanism by which pathogens interact with host autophagy remains mostly unclear. However, traditional strategies often have difficulty identifying host proteins that interact with effectors due to the weak, dynamic, and transient nature of these interactions. Here, we found that Enteropathogenic Escherichia coli (EPEC) regulates autophagosome formation in host cells dependent on effector NleE. The 26S Proteasome Regulatory Subunit 10 (PSMD10) was identified as a direct interaction partner of NleE in living cells by employing genetically incorporated crosslinkers. Pairwise chemical crosslinking revealed that NleE interacts with the N-terminus of PSMD10. We demonstrated that PSMD10 homodimerization is necessary for its interaction with ATG7 and promotion of autophagy, but not necessary for PSMD10 interaction with ATG12. Therefore, NleE-mediated PSMD10 in monomeric state attenuates host autophagosome formation. Our study reveals the mechanism through which EPEC attenuates host autophagy activity.