College of Veterinary Medicine, China Agricultural University, Beijing, China.
CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
mBio. 2019 Oct 29;10(5):e02483-19. doi: 10.1128/mBio.02483-19.
E165R, a highly specific dUTP nucleotidohydrolase (dUTPase) encoded by the African swine fever virus (ASFV) genome, is required for productive replication of ASFV in swine macrophages. Here, we solved the high-resolution crystal structures of E165R in its apo state and in complex with its product dUMP. Structural analysis explicitly defined the architecture of the active site of the enzyme as well as the interaction between the active site and the dUMP ligand. By comparing the ASFV E165R structure with dUTPase structures from other species, we found that the active site of E165R is highly similar to those of dUTPases from and , against which small-molecule chemicals have been developed, which could be the potential drug or lead compound candidates for ASFV. Our results provide important basis for anti-ASFV drug design by targeting E165R. African swine fever virus (ASFV), an affecting pigs and wild boars with up to 100% case fatality rate, is currently rampaging throughout China and some other countries in Asia. There is an urgent need to develop therapeutic and preventive reagents against the virus. Our crystallographic and biochemical studies reveal that ASFV E165R is a member of trimeric dUTP nucleotidohydrolase (dUTPase) family that catalyzes the hydrolysis of dUTP into dUMP. Our apo-E165R and E165R-dUMP structures reveal the constitutive residues and the configuration of the active center of this enzyme in rich detail and give evidence that the active center of E165R is very similar to that of dUTPases from and , which have already been used as targets for designing drugs. Therefore, our high-resolution structures of E165R provide useful structural information for chemotherapeutic drug design.
E165R 是非洲猪瘟病毒 (ASFV) 基因组编码的一种高度特异性的 dUTP 核苷水解酶 (dUTPase)，对于 ASFV 在猪巨噬细胞中的复制是必需的。在这里，我们解析了 E165R 的 apo 状态及其与产物 dUMP 复合物的高分辨率晶体结构。结构分析明确了酶的活性位点的结构以及活性位点与 dUMP 配体之间的相互作用。通过将 ASFV E165R 结构与来自其他物种的 dUTPase 结构进行比较，我们发现 E165R 的活性位点与来自 和 的 dUTPases 非常相似，针对这些 dUTPase，已经开发出了小分子化学物质，这些物质可能成为 ASFV 的潜在药物或先导化合物候选物。我们的研究结果为针对 E165R 的抗 ASFV 药物设计提供了重要依据。非洲猪瘟病毒 (ASFV) 是一种烈性传染病，感染猪和野猪后死亡率高达 100%，目前正在中国和亚洲其他一些国家肆虐。迫切需要开发针对该病毒的治疗和预防试剂。我们的晶体学和生物化学研究表明，ASFV E165R 是三聚体 dUTP 核苷水解酶 (dUTPase) 家族的成员，该酶催化 dUTP 水解为 dUMP。我们的 apo-E165R 和 E165R-dUMP 结构以丰富的细节揭示了该酶的组成性残基和活性中心的构象，并证明 E165R 的活性中心与 和 的 dUTPases 非常相似，这些酶已被用于设计药物的靶点。因此，我们的 E165R 高分辨率结构为化学治疗药物设计提供了有用的结构信息。
