Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, China.
Virulence. 2022 Dec;13(1):1849-1867. doi: 10.1080/21505594.2022.2136433.
Previously, we reported that the mediation of Newcastle disease virus (NDV) pathogenicity by the YLMY motif depends mainly on the regulation of F protein transport to the cell surface. The virus and host determinants that govern this intracellular trafficking remain unknown. Here, we confirmed that host adaptor protein (AP) complexes are involved in NDV infection using small interfering RNA. The transport of viral F protein to the cell surface depends on host transport proteins. We observed that the trends for host expression of AP complexes AP1M1 and AP2M1 were similar to those of mutated F proteins, especially in the membrane protein. NDV F protein interacted with AP1M1 and AP2M1, and the YLMY motif influenced this interaction. Knockdown of AP1M1 or AP2M1 suppressed the intracellular and extracellular virus titre of mutated-YLMY-motif NDVs, especially rSG10*-F/Y527A and rSG10*-F/Y524AY527A, to varying degrees. Therefore, the YLMY motif regulates AP-mediated viral F protein transportation from the cytoplasm to the cell surface and subsequently affects viral titer. We further found that the YLMY-motif mutants were differently associated with the process of AAK1 and GAK kinase-mediated AP - viral F protein interaction. These data demonstrate that the essential YLMY motif located in the NDV F protein cytoplasmic tail recruits AP to direct the F protein to the cell surface, which is necessary for its ability to affect virus budding. This study provides support for a deeper understanding of virus and host determinants that facilitate virus trafficking, which can be exploited in the design of novel antiviral therapies.
先前，我们报道了新城疫病毒（NDV）致病性的 YLMY 基序主要依赖于 F 蛋白向细胞表面运输的调节。控制这种细胞内运输的病毒和宿主决定因素尚不清楚。在这里，我们使用小干扰 RNA 证实了宿主衔接蛋白（AP）复合物参与了 NDV 感染。病毒 F 蛋白向细胞表面的转运依赖于宿主转运蛋白。我们观察到，AP 复合物 AP1M1 和 AP2M1 的宿主表达趋势与突变 F 蛋白相似，尤其是在膜蛋白中。NDV F 蛋白与 AP1M1 和 AP2M1 相互作用，并且 YLMY 基序影响这种相互作用。AP1M1 或 AP2M1 的敲低抑制了突变-YLMY 基序 NDV 的细胞内和细胞外病毒滴度，尤其是 rSG10*-F/Y527A 和 rSG10*-F/Y524AY527A，程度不同。因此，YLMY 基序调节 AP 介导的病毒 F 蛋白从细胞质到细胞表面的运输，进而影响病毒滴度。我们进一步发现，YLMY 基序突变体与 AAK1 和 GAK 激酶介导的 AP - 病毒 F 蛋白相互作用的过程不同相关。这些数据表明，位于 NDV F 蛋白胞质尾中的必需 YLMY 基序募集 AP 以将 F 蛋白引导至细胞表面，这对于其影响病毒出芽的能力是必要的。本研究为深入了解促进病毒运输的病毒和宿主决定因素提供了支持，这可用于设计新型抗病毒疗法。
