线粒体膜蛋白SLC25A23在I型干扰素抗病毒天然免疫中的调控作用及机制研究

SLC25A23, a kind of mitochondrial membrane transporter, belongs to the human solute carrier protein family 25A and member 23. The carrier imports adenine nucleotides into mitochondria matrix, taking an important role in the process of cell growth and energy metabolism. However, there is no study related to SLC25A23 and viral replication. Previously, we sought to identify the host-dependence factors by CRISPR GeCKO genome-wide screening library, and for the first time found that SLC25A23 was down regulated upon viral infection. Knock down of SLC25A23 could significantly inhibit vesicular stomatitis virus replication. Further experiments showed that SLC25A23 assisted viral replication by negative regulating type I interferon. Based on these evidences, we speculated that SLC25A23 can regulate type I interferon signaling pathways. In this study, we propose to further study of type I interferon regulation mechanism by gene overexpression or siRNA knockdown, the fluorescent microscopy, co-immune precipitation and other technical methods. These include the interaction of SLC25A23 with host proteins and molecular pathways downstream of transduction. This study will elucidate the new mechanism of SLC25A23 for type I interferon regulation, and also provide novel basis for potential targets in anti-viral drug design.

人溶质载体蛋白SLC25A23，是一种线粒体膜转运蛋白，其功能是将腺嘌呤核苷酸从细胞质转运至线粒体基质，在细胞能量代谢和生长过程中起重要作用，但是其与病毒复制还未见任何报道。前期我们通过CRISPR 全基因组文库筛选病毒复制宿主依赖因子，首次发现病毒感染过程中，SLC25A23表达减少。下调SLC25A23能够显著抑制水泡性性口炎病毒VSV复制，进一步实验表明SLC25A23通过负向调节I型干扰素产生辅助病毒复制，因此我们推测SLC25A23可调控I型干扰素产生，具有抗病毒功能。本研究我们拟采用基因过表达和siRNA干扰技术、荧光标记显微技术、免疫共沉淀等多种技术手段，深入研究SLC25A23在I型干扰素产生通路中的调控机制，包括鉴定与SLC25A23相互作用的蛋白以及上下游关键信号分子的分析。项目开展将为I型干扰素产生调控机制研究提供全新视角，也有望为我们发展抗病毒药物提供新的策略。

人溶质载体蛋白SLC25A23，是一种线粒体膜转运蛋白，其功能是将腺嘌呤核苷酸从粒体外转运至线粒体基质，并将磷酸盐转运至线粒体外，在细胞能量代谢和生长过程中起重要作用，但是其与病毒复制还未见任何报道。在本项目我们通过CRISPR GeCKO全基因组文库筛选病毒复制宿主依赖因子，首次发现病毒感染过程中，SLC25A23表达减少。下调SLC25A23能够显著抑制水泡性性口炎病毒VSV复制，进一步实验表明SLC25A23通过负向调节I型干扰素产生辅助病毒复制。我们通过蛋白相互作用鉴定出SLC25A23和RIG-I信号通路中关键接头分子MAVS相互作用，能改变MAVS K63的泛素化而影响MAVS寡聚化，进一步我们能够质谱测序得知 SLC25A23是通过E3泛素连接酶来调控MAVS的寡聚化。因此我们认为SLC25A23和Trim31的结合，阻碍了Trim31对MAVS的K63的泛素化，降低了MAVS由于K63泛素导致的寡聚化，负向调控I型干扰素产生，且我们在小鼠体内，利用siRNA干扰技术，同样也证明下调SLC25A23之后，具有显著的抗病毒作用。项目的开展为I型干扰素产生调控机制研究提供全新视角，也有望为我们发展抗病毒药物提供新的策略。