Lin Ying, Zhu Yun, Jing Ling, Chen Yongjun, Xiao Xia, Lei Xiaobo, Xie Zhengde
NHC Key Laboratory of System Biology of Pathogens and Christophe Merieux Laboratory, National Institute of Pathogen Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P.R. China.
Beijing Key Laboratory of Core Technologies for the Prevention and Treatment of Emerging Infectious Diseases in Children, Beijing Research Center for Respiratory Infectious Diseases, Beijing, China.
Life Sci Alliance. 2025 Jun 19;8(9). doi: 10.26508/lsa.202503320. Print 2025 Sep.
Human adenovirus B7 (HAdV-B7) is a significant respiratory pathogen in children, associated with substantial morbidity and mortality. Despite its clinical importance, the molecular mechanisms of HAdV-B7-host interaction remain poorly elucidated. In this study, we performed a high-throughput gain-of-function cDNA library screening and identified glycogen synthase kinase 3α (GSK3A) as a key proviral factor facilitating HAdV-B7 replication. The overexpression of GSK3A enhanced viral replication, whereas knockdown or knockout inhibited it. Furthermore, the kinase-active S21A mutant significantly augmented viral replication, whereas the kinase-inactive Y279A and K148A mutants of GSK3A failed to support it, highlighting the importance of its kinase activity on HAdV-B7 replication. Notably, phosphoproteomic and co-immunoprecipitation assays (co-IP) revealed that GSK3A phosphorylates viral L4-22K protein at S78 and S81 residues in a partially kinase-dependent manner. Using structure modeling, protein-protein docking, and truncation assays, we mapped the interaction between GSK3A's kinase domain and the 92-168 aa region of the viral L4-22K protein. In addition, GSK3A acts as a broad-spectrum proviral factor for respiratory HAdV, particularly for Species B, corresponding to a high similarity in L4-22K sequences. As a result, we identified GSK3A as a crucial proviral host factor for HAdV replication and provided a promising avenue for targeting GSK3A in the development of antiviral therapies against HAdV infections.
人腺病毒B7（HAdV - B7）是儿童重要的呼吸道病原体，与较高的发病率和死亡率相关。尽管其具有临床重要性，但HAdV - B7与宿主相互作用的分子机制仍不清楚。在本研究中，我们进行了高通量功能获得性cDNA文库筛选，并确定糖原合酶激酶3α（GSK3A）是促进HAdV - B7复制的关键病毒前体因子。GSK3A的过表达增强了病毒复制，而敲低或敲除则抑制了病毒复制。此外，激酶活性的S21A突变体显著增强了病毒复制，而GSK3A的激酶失活Y279A和K148A突变体则无法支持病毒复制，突出了其激酶活性对HAdV - B7复制的重要性。值得注意的是，磷酸化蛋白质组学和免疫共沉淀分析（co - IP）表明，GSK3A以部分激酶依赖性方式在S78和S81残基处磷酸化病毒L4 - 22K蛋白。通过结构建模、蛋白质 - 蛋白质对接和截短分析，我们绘制了GSK3A激酶结构域与病毒L4 - 22K蛋白92 - 168 aa区域之间的相互作用。此外，GSK3A作为呼吸道HAdV的广谱病毒前体因子，特别是对于B种病毒，这与L4 - 22K序列的高度相似性相对应。因此，我们确定GSK3A是HAdV复制的关键病毒前体宿主因子，并为在开发抗HAdV感染的抗病毒疗法中靶向GSK3A提供了一条有前景的途径。
