The lipid kinase PI4KB, which generates phosphatidylinositol 4-phosphate (PI4P), is a key enzyme in regulating membrane transport and is also hijacked by multiple picornaviruses to mediate viral replication. PI4KB can interact with multiple protein binding partners, which are differentially manipulated by picornaviruses to facilitate replication. The protein c10orf76 is a PI4KB-associated protein that increases PI4P levels at the Golgi and is essential for the viral replication of specific enteroviruses. We used hydrogen-deuterium exchange mass spectrometry to characterize the c10orf76-PI4KB complex and reveal that binding is mediated by the kinase linker of PI4KB, with formation of the heterodimeric complex modulated by PKA-dependent phosphorylation. Complex-disrupting mutations demonstrate that PI4KB is required for membrane recruitment of c10orf76 to the Golgi, and that an intact c10orf76-PI4KB complex is required for the replication of c10orf76-dependent enteroviruses. Intriguingly, c10orf76 also contributed to proper Arf1 activation at the Golgi, providing a putative mechanism for the c10orf76-dependent increase in PI4P levels at the Golgi.

中文翻译：

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c10orf76-PI4KB 复合物的表征及其对高尔基体 PI4P 水平和肠道病毒复制的必要性。

脂质激酶 PI4KB 可产生 4-磷酸磷脂酰肌醇 (PI4P)，是调节膜运输的关键酶，也被多种小核糖核酸病毒劫持以介导病毒复制。PI4KB 可以与多种蛋白质结合伴侣相互作用，这些蛋白质结合伴侣由小核糖核酸病毒进行差异性操纵以促进复制。蛋白质 c10orf76 是一种 PI4KB 相关蛋白，可增加高尔基体的 PI4P 水平，对于特定肠道病毒的病毒复制至关重要。我们使用氢-氘交换质谱法来表征 c10orf76-PI4KB 复合物，并揭示结合是由 PI4KB 的激酶接头介导的，并通过 PKA 依赖性磷酸化调节异二聚体复合物的形成。复合物破坏突变表明，PI4KB 是 c10orf76 膜募集至高尔基体所必需的，并且完整的 c10orf76-PI4KB 复合物是 c10orf76 依赖性肠道病毒复制所必需的。有趣的是，c10orf76 还有助于高尔基体中 Arf1 的适当激活，为高尔基体中 PI4P 水平的 c10orf76 依赖性增加提供了假定机制。