Chen Shuying, Yang Shanrong, Li Xiaoning, Xiang Junqi, Cai Jiangyu, Wang Yaokai, Li Qingqing, Zang Na, Wang Jiaxu, Shang Jian, Wan Yushun
College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
Henan Institute of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou 450052, China.
Viruses. 2025 Jul 22;17(8):1025. doi: 10.3390/v17081025.
Coronaviruses, particularly those classified as highly pathogenic species, pose a significant threat to global health. These viruses hijack host cellular membranes and proteins to facilitate their replication, primarily through the formation of replication organelles (ROs). However, the precise regulatory mechanisms underlying RO formation remain poorly understood. To elucidate these mechanisms, we conducted mass spectrometry analyses, identifying interactions between the host protein DnaJ homolog subfamily C member 11 (DNAJC11) and the SARS-CoV-2 nonstructural protein 3 (NSP3) protein. Notably, results showed that DNAJC11 depletion reduces SARS-CoV-2 infection, indicating possible positive regulatory involvement. But the ectopic expression of DNAJC11 did not lead to marked alterations in immune or inflammatory responses. DNAJC11 enhanced NSP3 expression stability through endogenous apoptosis pathways and facilitated its interaction with NSP4, thereby promoting the formation of double-membrane vesicles (DMVs). Knockdown of DNAJC11 reduced DMV number and size, accompanied by dysregulation of the endoplasmic reticulum and mitochondria. However, supplementation with DNAJC11 restored both DMV number and size. These findings provide novel insights into the role of DNAJC11 as a host factor that modulates DMV formation and supports SARS-CoV-2 replication by targeting the NSP3 protein. This study advances our understanding of the molecular interactions between host and viral components and highlights DNAJC11 as a potential target for antiviral interventions.
冠状病毒，尤其是那些被归类为高致病性的病毒，对全球健康构成重大威胁。这些病毒劫持宿主细胞膜和蛋白质以促进其复制，主要是通过形成复制细胞器（ROs）。然而，RO形成背后的确切调控机制仍知之甚少。为了阐明这些机制，我们进行了质谱分析，确定了宿主蛋白DnaJ同源亚家族C成员11（DNAJC11）与严重急性呼吸综合征冠状病毒2（SARS-CoV-2）非结构蛋白3（NSP3）之间的相互作用。值得注意的是，结果表明DNAJC11的缺失会降低SARS-CoV-2感染，表明可能存在正向调控作用。但是DNAJC11的异位表达并未导致免疫或炎症反应的明显改变。DNAJC11通过内源性凋亡途径增强NSP3表达稳定性，并促进其与NSP4的相互作用，从而促进双膜囊泡（DMVs）的形成。敲低DNAJC11会减少DMV的数量和大小，同时伴有内质网和线粒体的失调。然而，补充DNAJC11可恢复DMV的数量和大小。这些发现为DNAJC11作为宿主因子的作用提供了新的见解，该因子通过靶向NSP3蛋白来调节DMV形成并支持SARS-CoV-2复制。这项研究推进了我们对宿主与病毒成分之间分子相互作用的理解，并突出了DNAJC11作为抗病毒干预潜在靶点的地位。
