Phospho-proteomics identifies a critical role of ATF2 in pseudorabies virus replication  

作　　者：Fang-Fang Jiang Ren-Qi Wang Chao-Yue Guo Ke Zheng Hai-Long Liu Le Su Sheng-Song Xie Huan-Chun Chen Zheng-Fei Liu 

机构地区：[1]State Key Laboratory of Agricultural Microbiology,Hongshan Laboratory and Key laboratory of Preventive Veterinary Medicine in Hubei Province,College of Veterinary Medicine,Huazhong Agricultural University,Wuhan,430070,China [2]Key Lab of Agricultural Animal Genetics,Breeding,and Reproduction of Ministry of Education,Huazhong Agricultural University,Wuhan,430070,China

出　　处：《Virologica Sinica》2022年第4期591-600,共10页中国病毒学（英文版）

基　　金：the Natural Science Foundation of China(32170155,31770191);the Major Science and Technology Projects of Hubei Province(2021ABA005)to Z.F.L。

摘　　要：Pseudorabies virus(PRV),an etiological agent of pseudorabies in livestock,has negatively affected the porcine industry all over the world.Epithelial cells are reported as the first site of PRV infection.However,the role of host proteins and its related signaling pathways in PRV replication is largely unclear.In this study,we performed a quantitative phosphoproteomics screening on PRV-infected porcine kidney(PK-15)epithelial cells.Totally 5723phosphopeptides,corresponding to 2180 proteins,were obtained,and the phosphorylated states of 810 proteins were significantly different in PRV-infected cells compared with mock-infected cells(P<0.05).GO and KEGG analysis revealed that these differentially expressed phosphorylated proteins were predominantly related to RNA transport and MAPK signaling pathways.Further functional studies of NF-κB,transcription activator factor-2(ATF2),MAX and SOS genes in MAPK signaling pathway were analyzed using RNA interference(RNAi)knockdown.It showed that only ATF2-knockdown reduces both PRV titer and viral genome copy number.JNK pathway inhibition and CRISPR/Cas9 gene knockout showed that ATF2 was required for the effective replication of PRV,especially during the biogenesis of viral genome DNA.Subsequently,by overexpression of the ATF2 gene and point mutation of the amino acid positions 69/71 of ATF2,it was further demonstrated that the phosphorylation of ATF2 promoted PRV replication.These findings suggest that ATF2 may provide potential therapeutic target for inhibiting PRV infection.

关 键 词：Pseudorabies virus(PRV) Transcription activator factor-2(ATF2) ITRAQ PROTEOMICS PHOSPHORYLATION 

分 类 号：S852.65[农业科学—基础兽医学]