Crystal structure and biochemical activity of the macrodomain from rubella virus p150
Crystal structure and biochemical activity of the macrodomain from rubella virus p150
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DOI：
10.1128/jvi.01777-23
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发表时间：
2024-01-30
影响因子：
5.4
通讯作者：
Modis，Yorgo
中科院分区：
文献类型：
--
作者：
Stoll，Guido A.;Nikolopoulos，Nikos;Modis，Yorgo
风疹病毒编码一种具有RNA聚合酶、甲基转移酶和类木瓜蛋白酶活性的非结构多蛋白，以及一个功能未知的大结构域。大结构域结合ADP-核糖加合物，这是一种翻译后修饰，在宿主与病毒的冲突中发挥关键作用。一些大分子结构域还可以去除单-ADP-核糖加合物或降解多-ADP-核糖链。在这里，我们报道了风疹病毒非结构蛋白P150的大结构域的高分辨晶体结构，有和没有ADP-核糖结合。整个折叠结构与各种非病毒物种的大D-型大结构域最为相似。风疹病毒大结构域中与ADP-核糖配位的残基的特殊组成和结构与甲型病毒大结构域的组成和结构最为相似。等温量热分析表明风疹病毒大分子结构域在溶液中与ADP-核糖结合。酶分析表明，风疹病毒大分子结构域既能水解单-ADP核糖加合物，也能水解多-ADP-核糖加合物。定点突变鉴定了单腺苷二磷酸核糖水解酶活性所需的Asn39和Cys49。重要的是风疹病毒仍然是一个全球健康威胁。妊娠期风疹感染可导致严重的先天性病理，目前尚无抗病毒治疗方法。我们的工作表明，像α病毒和冠状病毒一样，RubiVirus编码一个结构保守的大结构域折叠的单ADP核糖水解酶，以抵消宿主先天性免疫反应中多聚ADP核糖聚合酶(PARP)的MAR化。我们的结构数据将指导未来开发针对风疹或相关病毒感染的新型抗病毒疗法的努力。
Rubella virus encodes a nonstructural polyprotein with RNA polymerase, methyltransferase, and papain-like cysteine protease activities, along with a putative macrodomain of unknown function. Macrodomains bind ADP–ribose adducts, a post-translational modification that plays a key role in host–virus conflicts. Some macrodomains can also remove the mono-ADP–ribose adduct or degrade poly-ADP–ribose chains. Here, we report high-resolution crystal structures of the macrodomain from rubella virus nonstructural protein p150, with and without ADP–ribose binding. The overall fold is most similar to macroD-type macrodomains from various nonviral species. The specific composition and structure of the residues that coordinate ADP–ribose in the rubella virus macrodomain are most similar to those of macrodomains from alphaviruses. Isothermal calorimetry shows that the rubella virus macrodomain binds ADP–ribose in solution. Enzyme assays show that the rubella virus macrodomain can hydrolyze both mono- and poly-ADP–ribose adducts. Site-directed mutagenesis identifies Asn39 and Cys49 required for mono-ADP-ribosylhydrolase (de-MARylation) activity.IMPORTANCERubella virus remains a global health threat. Rubella infections during pregnancy can cause serious congenital pathology, for which no antiviral treatments are available. Our work demonstrates that, like alpha- and coronaviruses, rubiviruses encode a mono-ADP-ribosylhydrolase with a structurally conserved macrodomain fold to counteract MARylation by poly (ADP–ribose) polymerases (PARPs) in the host innate immune response. Our structural data will guide future efforts to develop novel antiviral therapeutics against rubella or infections with related viruses.