Encephalomyocarditis virus leader is phosphorylated by CK2 and syk as a requirement for subsequent phosphorylation of cellular nucleoporins.

Abstract

Encephalomyocarditis virus and Theilovirus are species in the Cardiovirus genus of the Picornaviridae family. For all cardioviruses, the viral polyprotein is initiated with a short Leader (L) protein unique to this genus. The nuclear magnetic resonance (NMR) structure of LE from encephalomyocarditis virus (EMCV) has been determined. The protein has an NH2-proximal CHCC zinc finger, a central linker, and a contiguous, highly acidic motif. The theiloviruses encode the same domains, with one or two additional, COOH-proximal domains, characteristic of the human Saffold viruses (SafV) and Theiler's murine encephalomyelitis viruses (TMEV), respectively. The expression of a cardiovirus L, in recombinant form, or during infection/transfection, triggers an extensive, cell-dependent, antihost phosphorylation cascade, targeting nucleoporins (Nups) that form the hydrophobic core of nuclear pore complexes (NPC). The consequent inhibition of active nucleocytoplasmic trafficking is potent and prevents the host from mounting an effective antiviral response. For this inhibition, the L proteins themselves must be phosphorylated. In cells (extracts or recombinant form), LE was shown to be phosphorylated at Thr47 and Tyr41. The first reaction (Thr47), catalyzed by casein kinase 2 (CK2), is an obligatory precedent to the second event (Tyr41), catalyzed by spleen tyrosine kinase (Syk). Site mutations in LE, or kinase-specific inhibitors, prevented LE phosphorylation and subsequent Nup phosphorylation. Parallel experiments with LS (SafV-2) and LT (TMEV BeAn) proteins confirmed the general cardiovirus requirement for L phosphorylation, but CK2 was not the culpable kinase. It is likely that LS and LT are both activated by alternative kinases in different cell types, probably reactive within the Theilo-specific domains. IMPORTANCE An understanding of the diverse methods used by viruses to interfere with cellular processes is important because they can teach us how to control virus infections. This report shows how viruses in the same genus use different cellular enzymes to phosphorylate their proteins. If these processes are interfered with, the viruses are severely disabled.

• Authors: Holly A Basta, Valjean R Bacot-Davis, Jessica J Ciomperlik, Ann C Palmenberg
• Journal: Journal of virology (J Virol) Vol. 88 Issue 4 Pg. 2219-26 (Feb 2014) ISSN: 1098-5514 [Electronic] United States
• PMID: 24335301 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Intracellular Signaling Peptides and Proteins
• Nuclear Pore Complex Proteins
• Viral Proteins
• Protein-Tyrosine Kinases
• SYK protein, human
• Syk Kinase
• Casein Kinase II

Topics

• Base Sequence
• Blotting, Western
• Casein Kinase II (metabolism)
• Encephalomyocarditis virus (genetics)
• Intracellular Signaling Peptides and Proteins (metabolism)
• Magnetic Resonance Spectroscopy
• Molecular Sequence Data
• Mutagenesis, Site-Directed
• Nuclear Pore Complex Proteins (metabolism)
• Phosphorylation
• Plasmids (genetics)
• Protein-Tyrosine Kinases (metabolism)
• Sequence Alignment
• Sequence Analysis, DNA
• Syk Kinase
• Theilovirus (genetics)
• Viral Proteins (genetics, metabolism)