Proteolytic cleavage of the catalytic subunit of DNA-dependent protein kinase during poliovirus infection

Kareem L. Graham, Kurt E Gustin, Carlos Rivera, N. Muge Kuyumcu-Martinez, Sunny S. Choe, Richard E. Lloyd, Peter Sarnow, Paul J. Utz

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

DNA-dependent protein kinase (DNA-PK) is a serine/threonine kinase that has critical roles in DNA double-strand break repair, as well as B- and T-cell antigen receptor rearrangement. The DNA-PK enzyme consists of the Ku regulatory subunit and a 450-kDa catalytic subunit termed DNA-PKCS. Both of these subunits are autoantigens associated with connective tissue diseases such as systemic lupus erythematosus (SLE) and scleroderma. In this report, we show that DNA-PKCS is cleaved during poliovirus infection of HeLa cells. Cleavage was visible as early as 1.5 h postinfection (hpi) and resulted in an approximately 40% reduction in the levels of native protein by 5.5 hpi. Consistent with this observation, the activity of the DNA-PKCS enzyme was also reduced during viral infection, as determined by immunoprecipitation kinase assays. Although it has previously been shown that DNA-PKCS is a substrate of caspase-3 in vitro, the protein was still cleaved during poliovirus infection of the caspase-3-deficient MCF-7 cell line. Cleavage was not prevented by infection in the presence of a soluble caspase inhibitor, suggesting that cleavage in vivo was independent of host caspase activation. DNA-PKCS is directly cleaved by a picornaviral 2A protease in vitro, producing a fragment similar in size to the cleavage product observed in vivo. Taken together, our results indicate that DNA-PKCS is cleaved by the 2A protease during poliovirus infection. Proteolytic cleavage of DNA-PKCS during poliovirus infection may contribute to inhibition of host immune responses. Furthermore, cleavage of autoantigens by viral proteases may target these proteins for the autoimmune response by generating novel, or "immunocryptic," protein fragments.

Original languageEnglish (US)
Pages (from-to)6313-6321
Number of pages9
JournalJournal of Virology
Volume78
Issue number12
DOIs
StatePublished - Jun 2004
Externally publishedYes

Fingerprint

lysyl-arginyl-alanyl-lysyl-alanyl-lysyl-threonyl-threonyl-lysyl-lysyl-arginine
DNA-Activated Protein Kinase
Catalytic DNA
Enterovirus C
Poliovirus
protein subunits
protein kinases
Catalytic Domain
DNA
Infection
infection
Autoantigens
autoantigens
Caspase 3
proteinases
Proteins
Peptide Hydrolases
caspases
caspase-3
scleroderma (skin disease)

ASJC Scopus subject areas

  • Immunology

Cite this

Graham, K. L., Gustin, K. E., Rivera, C., Kuyumcu-Martinez, N. M., Choe, S. S., Lloyd, R. E., ... Utz, P. J. (2004). Proteolytic cleavage of the catalytic subunit of DNA-dependent protein kinase during poliovirus infection. Journal of Virology, 78(12), 6313-6321. https://doi.org/10.1128/JVI.78.12.6313-6321.2004

Proteolytic cleavage of the catalytic subunit of DNA-dependent protein kinase during poliovirus infection. / Graham, Kareem L.; Gustin, Kurt E; Rivera, Carlos; Kuyumcu-Martinez, N. Muge; Choe, Sunny S.; Lloyd, Richard E.; Sarnow, Peter; Utz, Paul J.

In: Journal of Virology, Vol. 78, No. 12, 06.2004, p. 6313-6321.

Research output: Contribution to journalArticle

Graham, KL, Gustin, KE, Rivera, C, Kuyumcu-Martinez, NM, Choe, SS, Lloyd, RE, Sarnow, P & Utz, PJ 2004, 'Proteolytic cleavage of the catalytic subunit of DNA-dependent protein kinase during poliovirus infection', Journal of Virology, vol. 78, no. 12, pp. 6313-6321. https://doi.org/10.1128/JVI.78.12.6313-6321.2004
Graham, Kareem L. ; Gustin, Kurt E ; Rivera, Carlos ; Kuyumcu-Martinez, N. Muge ; Choe, Sunny S. ; Lloyd, Richard E. ; Sarnow, Peter ; Utz, Paul J. / Proteolytic cleavage of the catalytic subunit of DNA-dependent protein kinase during poliovirus infection. In: Journal of Virology. 2004 ; Vol. 78, No. 12. pp. 6313-6321.
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