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.

Porcine Circovirus Type 2 Induces ORF3-Independent Mitochondrial Apoptosis via PERK Activation and Elevation of Cytosolic Calcium

“…Here we confirmed that ΔORF3 infection or Cap expression significantly induced cytosolic Ca 2ϩ and ROS levels and caspase-3 and -9 activities, accompanied by a loss of MMP, which was preventable by downregulation of PERK either by inhibition of PERK phosphorylation with GSK or by short hairpin PERK (shPERK). Chemical treatment could have diverse effects other than the expected impact, particularly with 4-PBA, which could arrest the cell cycle and prevent the entry into S phase, favoring PCV2 replication (35,36). To exclude the potential nonspecific effects of chemical treatment, we also used shRNA to specifically silence PERK expression.…”

Comparative transcriptome analysis reveals the role of p53 signalling pathway during red‐spotted grouper nervous necrosis virus infection in Lateolabrax japonicus brain cells

“…Recently, numbers of studies have shown that p53 signalling pathway is involved in the process of different virus infection and replication. For example, porcine circovirus type 2 infection induced S phase accumulation to promote viral replication in host cells via activation of the p53 signalling pathway (Xu et al, ). Classical swine fever virus Shimen regulated p53 signalling pathway to subvert host innate immune response (Ning et al, ).…”

Contribution of the swine model in the study of human sexually transmitted infections

“…Currently, this technology is used for molecular biology analyses in vitro, e.g. to study the role of apoptosis-inducing molecule p53 in an infection with the Porcine circovirus type 2 (Xu et al, 2016). In vivo, the CRISPR/Cas9 was A C C E P T E D M A N U S C R I P T 32 used to generate CD163 knockout pigs (Burkard et al, 2017).…”