蛋白激酶A通过磷酸化埃博拉病毒VP35调控病毒复制Protein Kinase A Mediated Phosphorylation of Ebola Virus VP35 Regulates the Viral Replication
靳京,张迅,王迪,汪婷婷,朱林,曹诚
摘要(Abstract)：
埃博拉病毒病（Ebola virus disease,EVD）是由埃博拉病毒（Ebola virus,EBOV）感染所引起的急性出血性伴多脏器损害的烈性传染病。VP35蛋白是EBOV RNA合成必需的聚合酶辅助因子，并可通过阻断干扰素途径抑制机体先天性免疫，其磷酸化可以促进病毒复制。我们前期研究发现EBOV VP35与A激酶相互作用蛋白1(A-kinase interacting protein 1,AKIP1)相互作用并激活蛋白激酶A(Protein kinase A,PKA)，但PKA是否可磷酸化VP35，从而调控EBOV复制尚不清楚。本研究发现，PKA激活剂Forskolin(FSK)及8-Bromo-cAMP促进VP35丝氨酸磷酸化，而PKA抑制剂H89及蛋白激酶抑制剂肽（Protein kinase inhibitor peptide,PKI）抑制VP35磷酸化。通过质谱分析鉴定出VP35存在多个磷酸化位点，VP35 S187A突变后，VP35磷酸化显著减弱，且PKA激活剂FSK和8-BromocAMP不能促进VP35 S187A的磷酸化，提示S187是PKA介导VP35磷酸化的主要位点。接着，利用可模拟病毒生命周期的EBOV trVLP系统研究了VP35磷酸化对病毒复制的影响，发现携带VP35 S187A突变的EBOV trVLP在细胞中的复制降低17倍，而H89和PKI处理并不能进一步抑制EBOV trVLP的增殖。这些结果表明，PKA通过介导VP35 S187位的磷酸化，促进EBOV trVLP的复制。此外，VP35 S187不影响VP35拮抗干扰素β产生的功能。本研究发现PKA通过磷酸化VP35 S187调控EBOV的复制，提示PKA抑制剂可以用于拮抗EBOV的增殖，进而为EVD的治疗提供新思路。
关键词(KeyWords)： 蛋白激酶A;埃博拉病毒;VP35磷酸化;VP35 S187;病毒复制
基金项目(Foundation): 中国科学院武汉国家生物安全实验室高端用户培育项目（项目号：2022ACCP-MS04）,题目：埃博拉病毒复制分子机制研究及亨尼帕病毒/埃博拉病毒中和抗体评价;; 国家科技重大专项（项目号：2018ZX09711003-005-005）,题目：烈性呼吸道及出血热反向遗传学研究及药效学评价~~
作者(Author): 靳京,张迅,王迪,汪婷婷,朱林,曹诚
DOI: 10.13242/j.cnki.bingduxuebao.004356
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