Coronavirus Infection Induces DNA Replication Stress Partly through Interaction of Its Nonstructural Protein 13 with the p125 Subunit of DNA Polymerase δ

• Published in: The Journal of biological chemistry Vol. 286; no. 45; pp. 39546 - 39559
• Main Authors: Xu, Ling Hui, Huang, Mei, Fang, Shou Guo, Liu, Ding Xiang
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 11.11.2011; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Ataxia Telangiectasia Mutated Proteins; Cell Cycle Checkpoints; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Chlorocebus aethiops; Coronavirus; Coronavirus Infections - genetics; Coronavirus Infections - metabolism; DNA Damage; DNA Damage - genetics; DNA Polymerase; DNA Polymerase III - genetics; DNA Polymerase III - metabolism; DNA Polymerase δ; DNA Replication; DNA Replication Stress; HeLa Cells; Humans; Infectious bronchitis virus - physiology; Interaction; Interphase - genetics; Microbiology; nsp13; Protein Serine-Threonine Kinases - genetics; Protein Serine-Threonine Kinases - metabolism; Vero Cells; Viral Nonstructural Proteins - genetics; Viral Nonstructural Proteins - metabolism; Viral Protein; Viral Replication; Virus; Virus Replication - physiology; Virus; DNA Polymerase; Viral Replication; Viral Protein; Coronavirus; DNA Polymerase δ; DNA Replication Stress; Interaction; nsp13; DNA Damage
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M111.242206

Perturbation of cell cycle regulation is a characteristic feature of infection by many DNA and RNA viruses, including Coronavirus infectious bronchitis virus (IBV). IBV infection was shown to induce cell cycle arrest at both S and G2/M phases for the enhancement of viral replication and progeny production. However, the underlying mechanisms are not well explored. In this study we show that activation of cellular DNA damage response is one of the mechanisms exploited by Coronavirus to induce cell cycle arrest. An ATR-dependent cellular DNA damage response was shown to be activated by IBV infection. Suppression of the ATR kinase activity by chemical inhibitors and siRNA-mediated knockdown of ATR reduced the IBV-induced ATR signaling and inhibited the replication of IBV. Furthermore, yeast two-hybrid screens and subsequent biochemical and functional studies demonstrated that interaction between Coronavirus nsp13 and DNA polymerase δ induced DNA replication stress in IBV-infected cells. These findings indicate that the ATR signaling activated by IBV replication contributes to the IBV-induced S-phase arrest and is required for efficient IBV replication and progeny production.