TRIM7/RNF90 promotes autophagy via regulation of ATG7 ubiquitination during L. monocytogenes infection

Jie Wang; Xiao Qin; Yulu Huang; Qunmei Zhang; Jinyong Pei; Yi Wang; Idan Goren; Shujun Ma; Zhishan Song; Yanzi Liu; Hongxia Xing; Hui Wang; Bo Yang

doi:10.1080/15548627.2022.2162706

TRIM7/RNF90 promotes autophagy via regulation of ATG7 ubiquitination during L. monocytogenes infection

Autophagy. 2023 Jun;19(6):1844-1862. doi: 10.1080/15548627.2022.2162706. Epub 2023 Jan 1.

Authors

Jie Wang^{1

2}, Xiao Qin^{1

2}, Yulu Huang^{1

2}, Qunmei Zhang³, Jinyong Pei^{1

2}, Yi Wang^{1

2}, Idan Goren^{4

5}, Shujun Ma^{1

2}, Zhishan Song^{1

2}, Yanzi Liu⁶, Hongxia Xing⁷, Hui Wang^{1

2}, Bo Yang^{1

2}

Affiliations

¹ Henan Key Laboratory of Immunology and Targeted Drug, Xinxiang Medical University, Xinxiang, Henan, China.
² Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang, Henan, China.
³ Clinical Laboratory, The First Affiliated Hospital of Xinxiang Medical University, Weihui, County, China.
⁴ Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
⁵ Department of Inflammation and Immunity, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, USA.
⁶ Department of Laboratory Medicine, the Third Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan, China.
⁷ Xinxiang Key Laboratory of Movement Disorders, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan, China.

Abstract

L. monocytogenes is a widely used infection model for the research on pathogenesis and host defense against gram-positive intracellular bacteria. Emerging evidence indicates that posttranslational modifications play a critical role in the regulation of macroautophagy/autophagy. However, little is known about the posttranslational modifications of ATG7, the essential protein in the autophagy process. In this study, we demonstrated that the RING-type E3 ligase TRIM7/RNF90 positively regulated autophagosome accumulation by promoting the ubiquitination of ATG7 at K413, thereby affecting L. monocytogenes infection. TRIM7 expression was induced by a variety range of conditions, including starvation, rapamycin stimulation, and L. monocytogenes infection. TRIM7 deficiency in mice or cells resulted in elevated innate immune responses and increased L. monocytogenes infection. ATG7 was associated with TRIM7 and the positive regulatory role of TRIM7 in L. monocytogenes infection-, starvation- or rapamycin-induced autophagosome accumulation was suggested by TRIM7 deficiency, TRIM7 overexpression, and TRIM7 knockdown. Further mechanistic investigation indicated that TRIM7 promoted the K63-linked ubiquitination of ATG7 at K413 and ubiquitination at this site was required for the function of ATG7 in autophagy and L. monocytogenes infection. Thus, our findings suggested a new regulator in intracellular bacterial infection and autophagy, with a novel posttranslational modification targeting ATG7. This research may expand our understanding of host anti-bacterial defense and the role of autophagy in intracellular bacterial infection.Abbreviations: ATG3: autophagy related 3; ATG5: autophagy related 5; ATG7: autophagy related 7; ATG10: autophagy related 10; ATG12: autophagy related 12; ATG16L1: autophagy related 16 like 1; Baf A1: bafilomycin A₁; CQ: chloroquine; BMDC: bone marrow-derived dendritic cell; BMDM: bone marrow-derived macrophage; CFUs: colony-forming units; CXCL10/IP-10: C-X-C motif chemokine ligand 10; EBSS: Earle's balanced salt solution; ELISA: enzyme-linked immunosorbent assay; IFIT1/ISG56: interferon induced protein with tetratricopeptide repeats 1; IFNB/IFN-β: interferon beta; IL6: interleukin 6; IRF3, interferon regulatory factor 3; Lm: L. monocytogenes; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MEF: mouse embryonic fibroblast; MOI: multiplicity of infection; PLA: proximity ligation assay; PMA: phorbol myristate acetate; PMA-THP1, PMA-differentiated THP1; PMs: peritoneal macrophages; PTMs: posttranslational modifications; STING1, stimulator of interferon response cGAMP interactor 1; TBK1, TANK binding kinase 1; TNF/TNF-α: tumor necrosis factor; TRIM7/RNF90: tripartite motif containing; Hainan Provincial Natural Science Foundation of China.

Keywords: Autophagosome accumulation; E3 ligase; autophagy-related genes; intracellular bacteria; posttranslational modification; ring finger proteins.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Autophagy* / physiology
Fibroblasts*
Interferons
Mice
Transcription Factors
Ubiquitination

Substances

Transcription Factors
Interferons

Grants and funding

This work was supported by the National Natural Science Foundation of China Grants (U1704183, 31970847, 32070949, U2004103, and 32170871); Natural Science Foundation of Henan Province (212300410065, 222300420064) Hainan Provincial Natural Science Foundation of China and Program for Science & Technology Innovation Talents in Higher Education of Henan Province (23HASTIT050) Higher Education of Henan province.