TRIM7/RNF90 promotes autophagy via regulation of ATG7 ubiquitination during L. monocytogenes infection
TRIM7/RNF90 promotes autophagy via regulation of ATG7 ubiquitination during L. monocytogenes infection
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DOI：
10.1080/15548627.2022.2162706
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发表时间：
2022-12-31
期刊：
影响因子：
13.3
通讯作者：
Yang，Bo
中科院分区：
文献类型：
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作者：
Wang，Jie;Qin，Xiao;Yang，Bo
L.单核细胞生成是研究革兰氏阳性细胞内细菌致病机理和宿主防御的一种广泛使用的感染模型。新出现的证据表明，翻译后修饰在宏观自噬/自噬的调节中起着关键作用。然而，很少有人知道的翻译后修饰的ATG 7，在自噬过程中的基本蛋白质。在这项研究中，我们证明了RING型E3连接酶TRIM 7/RNF 90通过促进K413处ATG 7的泛素化来正向调节自噬体积累，从而影响L。单核细胞增多症TRIM 7的表达被多种条件诱导，包括饥饿、雷帕霉素刺激和L.单核细胞增多症小鼠或细胞中的TRIM 7缺陷导致先天性免疫应答升高和L.单核细胞增多症ATG 7与TRIM 7相关，TRIM 7在L中的正调控作用。TRIM 7缺陷、TRIM 7过表达和TRIM 7敲低提示单核细胞增生感染、饥饿或雷帕霉素诱导的自噬体积累。进一步的机制研究表明，TRIM 7促进了K63连接的K413处的ATG 7泛素化，该位点的泛素化是ATG 7在自噬和L中发挥功能所必需的。单核细胞增多症因此，我们的研究结果表明，细胞内细菌感染和自噬中的一种新的调节剂，具有靶向ATG 7的新型翻译后修饰。本研究可能扩大我们对宿主抗细菌防御和自噬在细胞内细菌感染中的作用的理解。缩写：ATG 3：自噬相关3; ATG 5：自噬相关5; ATG 7：自噬相关7; ATG 10：自噬相关10; ATG 12：自噬相关12; ATG 16 L1：自噬相关16 like 1; Baf A1：巴弗洛霉素A1; CQ：氯喹; BMDC：骨髓源性树突状细胞; BMDM：骨髓源性巨噬细胞; CFUs：集落形成单位; CXCL 10/IP-10：C-X-C基序趋化因子配体10; EBSS：Earle平衡盐溶液; ELISA：酶联免疫吸附测定; IFIT 1/ISG 56：干扰素诱导的四肽重复序列1蛋白; IFNB/IFN-β：干扰素β; IL 6：白细胞介素6; IRF 3，干扰素调节因子3;单核细胞增多症; MAP1LC 3/LC 3：微管相关蛋白1轻链3; MEF：小鼠胚胎成纤维细胞; MOI：感染复数; PLA：邻位连接试验; PMA：佛波醇肉豆蔻酸酯乙酸酯; PMA-THP 1，PMA分化的THP 1; PM：腹腔巨噬细胞; PTM：翻译后修饰; STING 1，干扰素应答刺激因子cGAMP相互作用因子1; TBK 1，TANK结合激酶1; TNF/TNF-α：肿瘤坏死因子; TRIM 7/RNF 90：含三重基序;海南省自然科学基金。
L. monocytogenesis 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 affectingL. monocytogenesinfection. TRIM7 expression was induced by a variety range of conditions, including starvation, rapamycin stimulation, andL. monocytogenesinfection. TRIM7 deficiency in mice or cells resulted in elevated innate immune responses and increasedL. monocytogenesinfection. ATG7 was associated with TRIM7 and the positive regulatory role of TRIM7 inL. monocytogenesinfection-, 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 andL. monocytogenesinfection. 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.