Impaired STING Pathway in Human Osteosarcoma U2OS Cells Contributes to the Growth of ICP0-Null Mutant Herpes Simplex Virus.
Human herpes simplex virus 1 (HSV-1) is a widespread pathogen, with 80% of the population being latently infected. To successfully evade the host, the virus has evolved strategies to counteract antiviral responses, including the gene-silencing and innate immunity machineries. The immediately early protein of the virus, infected cell protein 0 (ICP0), plays a central role in these processes. ICP0 blocks innate immunity, and one mechanism is by degrading hostile factors with its intrinsic E3 ligase activity. ICP0 also functions as a promiscuous transactivator, and it blocks repressor complexes to enable viral gene transcription. For these reasons, the growth of a ΔICP0 virus is impaired in most cells, except cells of the human osteosarcoma cell line U2OS, and it is only partially impaired in cells of the human osteosarcoma cell line Saos-2. We found that the two human osteosarcoma cell lines that supported the growth of the ΔICP0 virus failed to activate innate immune responses upon treatment with 2'3'-cyclic GAMP (2'3'-cGAMP), the natural agonist of STING (i.e., stimulator of interferon genes) or after infection with the ΔICP0 mutant virus. Innate immune responses were restored in these cells by transient expression of the STING protein but not after overexpression of interferon-inducible protein 16 (IFI16). Restoration of STING expression resulted in suppression of ΔICP0 virus gene expression and a decrease in viral yields. Overexpression of IFI16 also suppressed ΔICP0 virus gene expression, albeit to a lesser extent than STING. These data suggest that the susceptibility of U2OS and Saos-2 cells to the ΔICP0 HSV-1 is in part due to an impaired STING pathway.IMPORTANCEThe DNA sensor STING plays pivotal role in controlling HSV-1 infection both in cell culture and in mice. The HSV-1 genome encodes numerous proteins that are dedicated to combat host antiviral responses. The immediate early protein of the virus ICP0 plays major role in this process as it targets hostile host proteins for degradation with its E3 ligase activity, and it disrupts repressor complexes via protein-protein interaction to enable viral gene transcription. Therefore, the ΔICP0 HSV-1 virus is defective for growth in most cells, except the human osteosarcoma cell lines U2OS and Saos-2. We found that both cell lines that support ΔICP0 virus infection have defects in the STING DNA-sensing pathway, which partially accounts for the rescue of the ΔICP0 virus growth. Restoration of STING expression in these cells rescued innate immunity and suppressed ΔICP0 virus infection. This study underscores the importance of STING in the control of HSV-1.
University of Kansas Medical Center, Department of Microbiology, Molecular Genetics and Immunology, Kansas City, Kansas, USA.
Thibaut
T
Deschamps
University of Kansas Medical Center, Department of Microbiology, Molecular Genetics and Immunology, Kansas City, Kansas, USA mkalamvoki@kumc.edu.
Maria
M
Kalamvoki
D001859:Bone Neoplasms / Q000235:genetics / Q000473:pathology* / Q000821:virology; D045744:Cell Line, Tumor; D006152:Cyclic GMP / Q000494:pharmacology; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics*; D007113:Immunity, Innate / Q000235:genetics / Q000276:immunology; D008565:Membrane Proteins / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D012516:Osteosarcoma / Q000235:genetics / Q000473:pathology* / Q000821:virology; D010750:Phosphoproteins / Q000378:metabolism; D018139:Simplexvirus / Q000235:genetics / Q000254:growth & development / Q000276:immunology; D015534:Trans-Activators / Q000235:genetics; D044767:Ubiquitin-Protein Ligases / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics; D000099197:STING Protein
D006152:Cyclic GMP; D017874:Immediate-Early Proteins; D008565:Membrane Proteins; D009687:Nuclear Proteins; D010750:Phosphoproteins; D015534:Trans-Activators; D044767:Ubiquitin-Protein Ligases; C532857:STING1 protein, human; D000099197:STING Protein; C076700:IFI16 protein, human; C051804:Vmw110 protein, Human herpesvirus 1
ICP0; IFI16; STING; herpes simplex virus; innate immunity