title: p53 signaling modulation of cell cycle arrest and viral replication in porcine circovirus type 2 infection cells.
abstract: Porcine circovirus type 2 (PCV2) is a ubiquitous pathogen in the swine industry worldwide. Previous studies have shown that PCV2 infection induces host cell apoptosis through up-regulation of p53. To further identify the regulatory roles of p53 signaling in the process of PCV2 infection, we established p53 gene knockout PK15 cell lines using the genomic editor tool CRISPR/Cas9, and further investigated the roles of p53 in modulating the cell cycle and viral replication in this study. The results show that PCV2 infection induced obvious S phase accumulation in wild-type PK15 cells and a compromised S phase accumulation in the p53 gene mutation cells (813PK15p53m/m), but did not induce obvious S phase accumulation in the p53 gene knockout cells (148PK15p53-/-) compared with the respective mock infection. PCV2 infection activated p53 signaling, up-regulated the expression of p21, Cyclin E, and down-regulated Cyclin A, CDK2. In p53 deficient cells, however, PCV2-induced changes in Cyclin A, CDK2, and Cyclin E were efficiently reversed to the basal levels. Detection of PCV2 replication showed decreased viral ORF1 genomic DNA in p53 deficient cells (148PK15p533-/-) and p53 mutated cells (813PK15p53m/m) compared with p53 wild-type cells after different synchronization treatment. Furthermore, PCV2 viral genomic DNA and Cap protein levels were higher in the cells released from S phase synchronized cells than in the cells released from the G0/G1 phase or G2/M phase-synchronized, or asynchronous cells after 18 h post-infection. Taken together, this study demonstrates that PCV2 infection induces S phase accumulation to favor viral replication in host cells through activation of the p53 pathway.
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 22 Xinong Rd, Yangling, Shaanxi, 712100, People's Republic of China.
authors.forename: Dan
authors.initials: D
authors.lastname: Xu
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 22 Xinong Rd, Yangling, Shaanxi, 712100, People's Republic of China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Du
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 22 Xinong Rd, Yangling, Shaanxi, 712100, People's Republic of China.
authors.forename: Cong
authors.initials: C
authors.lastname: Han
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 22 Xinong Rd, Yangling, Shaanxi, 712100, People's Republic of China.
authors.forename: Zengguo
authors.initials: Z
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 22 Xinong Rd, Yangling, Shaanxi, 712100, People's Republic of China.
authors.forename: Xiujuan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 22 Xinong Rd, Yangling, Shaanxi, 712100, People's Republic of China.
authors.forename: Tongtong
authors.initials: T
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 22 Xinong Rd, Yangling, Shaanxi, 712100, People's Republic of China.
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Zhao
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 22 Xinong Rd, Yangling, Shaanxi, 712100, People's Republic of China. yonghuang@nwsuaf.edu.cn.
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 22 Xinong Rd, Yangling, Shaanxi, 712100, People's Republic of China. dwtong@nwsuaf.edu.cn.
authors.forename: Dewen
authors.initials: D
authors.lastname: Tong
mesh_terms: D000818:Animals; D015153:Blotting, Western; D064113:CRISPR-Cas Systems; D002453:Cell Cycle / Q000502:physiology*; D018173:Circoviridae Infections / Q000662:veterinary* / Q000821:virology; D017925:Circovirus / Q000502:physiology*; D019925:Cyclin A / Q000378:metabolism; D055785:Gene Knockdown Techniques; D015398:Signal Transduction / Q000502:physiology; D013552:Swine; D013553:Swine Diseases / Q000821:virology*; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*; D014779:Virus Replication / Q000502:physiology
chemical_list: D019925:Cyclin A; D016159:Tumor Suppressor Protein p53