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Journal of Virology, May 2000, p. 3975-3983, Vol. 74, No. 9
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

The Viral Nucleocapsid Protein of Transmissible Gastroenteritis Coronavirus (TGEV) Is Cleaved by Caspase-6 and -7 during TGEV-Induced Apoptosis

Jean-François Eléouët,1,* Elizabeth A. Slee,2,dagger Françoise Saurini,1 Nathalie Castagné,1 Didier Poncet,1 Carmen Garrido,3 Eric Solary,3 and Seamus J. Martin2,dagger

Unité de Virologie et Immunologie Moléculaires, Institut National de la Recherche Agronomique, 78350 Jouy-en-Josas,1 and U.F.R. Médecine et Pharmacie, INSERM U517, 21000 Dijon,3 France, and Molecular Cell Biology Laboratory, National University of Ireland, Maynooth, County Kildare, Ireland2

Received 15 November 1999/Accepted 31 January 2000

The transmissible gastroenteritis coronavirus (TGEV), like many other viruses, exerts much of its cytopathic effect through the induction of apoptosis of its host cell. Apoptosis is coordinated by a family of cysteine proteases, called caspases, that are activated during apoptosis and participate in dismantling the cell by cleaving key structural and regulatory proteins. We have explored the caspase activation events that are initiated upon infection of the human rectal tumor cell line HRT18 with TGEV. We show that TGEV infection results in the activation of caspase-3, -6, -7, -8, and -9 and cleavage of the caspase substrates eIF4GI, gelsolin, and alpha -fodrin. Surprisingly, the TGEV nucleoprotein (N) underwent proteolysis in parallel with the activation of caspases within the host cell. Cleavage of the N protein was inhibited by cell-permeative caspase inhibitors, suggesting that this viral structural protein is a target for host cell caspases. We show that the TGEV nucleoprotein is a substrate for both caspase-6 and -7, and using site-directed mutagenesis, we have mapped the cleavage site to VVPD359down-arrow . These data demonstrate that viral proteins can be targeted for destruction by the host cell death machinery.

* Corresponding author. Mailing address: INRA, Unité de Virologie et Immunologie Moléculaires, 78352 Jouy-en-Josas Cedex, France. Phone: (33) 1 34 65 26 41. Fax: (33) 1 34 65 26 21. E-mail: eleouet{at}biotec.jouy.inra.fr.

dagger Present address: Division of Molecular and Cell Biology, The Smurfit Institute of Genetics, Trinity College, Dublin 2, Ireland.

Journal of Virology, May 2000, p. 3975-3983, Vol. 74, No. 9
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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