Yang Bo, Shen Mengyang, Lu Chen, Wang Yi, Zhao Xin, Zhang Qunmei, Qin Xiao, Pei Jinyong, Wang Hui, Wang Jie
Xinxiang Key Laboratory of Inflammation and Immunology, School of Medical Technology, Xinxiang Medical University, Xinxiang, Henan, China.
Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, School of Medical Technology, Xinxiang Medical University, Xinxiang, Henan, China.
Autophagy. 2025 Apr;21(4):789-806. doi: 10.1080/15548627.2024.2429380. Epub 2024 Nov 28.
is widely used in the laboratory as an infection model for the research on pathogenesis and host defense against gram-positive intracellular bacteria. Macroautophagy (called simply "autophagy" hereafter), is important in the host defense against pathogens, such as bacteria, viruses, and parasites. BECN1 plays a pivotal role in the initiation of autophagy and accumulating evidence indicates that post-translational modifications of BECN1 provide multiple strategies for autophagy regulation. In this study, we demonstrated that the RING1-IBR-RING2 (RBR) family member RNF144A (ring finger protein 144A), which was induced by infection, promoted infection in an autophagy-dependent but STING1-independent pattern. deficiency in mice protected mice from infection with inhibited innate immune responses. Interestingly, RNF144A decreased -induced autophagosome accumulation. Mechanistic investigation indicated that RNF144A interacted with BECN1 and promoted its K48-linked ubiquitination, leading to the subsequent proteasome-dependent degradation of BECN1 and reduced autophagosome accumulation. Further study demonstrated that RNF144A promoted the ubiquitination of BECN1 at K117 and K427, and these two ubiquitination sites were essential to the role of BECN1 in autophagy and Lm infection. Thus, our findings suggested a new regulator in intracellular bacterial infection and autophagy, which may contribute to our understanding of host defense against intracellular bacterial infection via autophagy.: 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 A: bafilomycin A; BECN1: beclin 1; BMDC: bone marrow-derived dendritic cell; BMDM: bone marrow-derived macrophage; CFUs: colony-forming units; CHX: cycloheximide; CQ: chloroquine; 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: ; 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; RBR: RING1-IBR-RING2; RNF144A: ring finger protein 144A; STING1, stimulator of interferon response cGAMP interactor 1; TBK1, TANK binding kinase 1; TNF/TNF-α: tumor necrosis factor.
在实验室中被广泛用作研究革兰氏阳性细胞内细菌发病机制和宿主防御的感染模型。巨自噬（以下简称为“自噬”）在宿主抵御病原体（如细菌、病毒和寄生虫）的过程中很重要。BECN1在自噬起始中起关键作用，越来越多的证据表明BECN1的翻译后修饰为自噬调节提供了多种策略。在本研究中，我们证明了由感染诱导的RING1-IBR-RING2（RBR）家族成员RNF144A（环指蛋白144A）以自噬依赖性但不依赖STING1的模式促进感染。小鼠体内RNF144A缺陷通过抑制先天免疫反应保护小鼠免受感染。有趣的是，RNF144A减少了感染诱导的自噬体积累。机制研究表明，RNF144A与BECN1相互作用并促进其K48连接的泛素化，导致随后BECN1依赖蛋白酶体的降解和自噬体积累减少。进一步研究表明，RNF144A促进BECN1在K117和K427处的泛素化，这两个泛素化位点对于BECN1在自噬和单核细胞增多性李斯特菌感染中的作用至关重要。因此，我们的研究结果提示了细胞内细菌感染和自噬中的一种新调节因子，这可能有助于我们理解宿主通过自噬抵御细胞内细菌感染的机制。：ATG3：自噬相关3；ATG5：自噬相关5；ATG7：自噬相关7；ATG10：自噬相关10；ATG12：自噬相关12；ATG16L1：自噬相关16样蛋白1；巴弗洛霉素A；BECN1：Beclin 1；骨髓来源的树突状细胞；骨髓来源的巨噬细胞；集落形成单位；放线菌酮；氯喹；C-X-C基序趋化因子配体10；Earle平衡盐溶液；酶联免疫吸附测定；干扰素诱导的含四肽重复序列蛋白1；干扰素β；白细胞介素6；干扰素调节因子3；单核细胞增多性李斯特菌；微管相关蛋白1轻链3；小鼠胚胎成纤维细胞；感染复数；邻近连接分析；佛波酯；佛波酯分化的THP1；腹膜巨噬细胞；翻译后修饰；RBR：RING1-IBR-RING2；RNF144A：环指蛋白144A；干扰素反应cGAMP相互作用分子1；TANK结合激酶1；肿瘤坏死因子
