Genetically incorporated crosslinkers reveal NleE attenuates host autophagy dependent on PSMD10.

Jingxiang Li,Pan Li,Li Gao,Qi Sun,Haiyan Ren,Fangni Chai,Shiqian Qi,Shupan Guo,Li Zhou,Xiaoxiao Ouyang,Lunzhi Dai,Kefeng Lu,Wei Cheng,Zhihui Zhou

doi:10.7554/elife.69047

Jingxiang Li, Pan Li + Show 12 more

Open Access

https://doi.org/10.7554/elife.69047

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Journal: eLife	Publication Date: Jul 13, 2021
Citations: 5	License type: CC BY 4.0

Affiliation: West China Hospital of Sichuan University

Abstract

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.

Highlights

Autophagy, a process referring to engulfment of a portion of the cytosol in a double-membrane autophagosome to lysosomes for degradation, plays a vital role in the host response to pathogens
We found that LPS-induced autophagy was partially suppressed by the T3SS effector NleE (Figure 1A,B)
We examined whether NleE affects another LC3-mediated process

Summary

Introduction

A process referring to engulfment of a portion of the cytosol in a double-membrane autophagosome to lysosomes for degradation, plays a vital role in the host response to pathogens. Genetically encoded crosslinking technology has emerged as an attractive strategy for the investigation of native protein– protein interactions in living cells (Coin, 2018; Coin et al, 2013; Yang et al, 2016; Zhang et al, 2011; Zhang et al, 2017; Tang et al, 2018). This technology has been applied mostly to map peptide–protein and protein–protein interactions. NleE interacts with the N-terminus of PSMD10 and suppresses PSMD10 homodimerization

Results

Discussion

Materials and methods