title: Cleavage of DAP5 by coxsackievirus B3 2A protease facilitates viral replication and enhances apoptosis by altering translation of IRES-containing genes.
abstract: Cleavage of eukaryotic translation initiation factor 4G (eIF4G) by enterovirus proteases during infection leads to the shutoff of cellular cap-dependent translation, but does not affect the initiation of cap-independent translation of mRNAs containing an internal ribosome entry site (IRES). Death-associated protein 5 (DAP5), a structural homolog of eIF4G, is a translation initiation factor specific for IRES-containing mRNAs. Coxsackievirus B3 (CVB3) is a positive single-stranded RNA virus and a primary causal agent of human myocarditis. Its RNA genome harbors an IRES within the 5'-untranslated region and is translated by a cap-independent, IRES-driven mechanism. Previously, we have shown that DAP5 is cleaved during CVB3 infection. However, the protease responsible for cleavage, cleavage site and effects on the translation of target genes during CVB3 infection have not been investigated. In the present study, we demonstrated that viral protease 2A but not 3C is responsible for DAP5 cleavage, generating 45- and 52-kDa N- (DAP5-N) and C-terminal (DAP5-C) fragments, respectively. By site-directed mutagenesis, we found that DAP5 is cleaved at amino acid G434. Upon cleavage, DAP5-N largely translocated to the nucleus at the later time points of infection, whereas the DAP5-C largely remained in the cytoplasm. Overexpression of these DAP5 truncates demonstrated that DAP5-N retained the capability of initiating IRES-driven translation of apoptosis-associated p53, but not the prosurvival Bcl-2 (B-cell lymphoma 2) when compared with the full-length DAP5. Similarly, DAP5-N expression promoted CVB3 replication and progeny release; on the other hand, DAP5-C exerted a dominant-negative effect on cap-dependent translation. Taken together, viral protease 2A-mediated cleavage of DAP5 results in the production of two truncates that exert differential effects on protein translation of the IRES-containing genes, leading to enhanced host cell death.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.|The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: P J
authors.initials: PJ
authors.lastname: Hanson
authors.affiliation: Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.|The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: X
authors.initials: X
authors.lastname: Ye
authors.affiliation: Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.|The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: Y
authors.initials: Y
authors.lastname: Qiu
authors.affiliation: Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.|The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: H M
authors.initials: HM
authors.lastname: Zhang
authors.affiliation: Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.|The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: M G
authors.initials: MG
authors.lastname: Hemida
authors.affiliation: The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: F
authors.initials: F
authors.lastname: Wang
authors.affiliation: Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.|The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: T
authors.initials: T
authors.lastname: Lim
authors.affiliation: The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Gu
authors.affiliation: The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: B
authors.initials: B
authors.lastname: Cho
authors.affiliation: The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: H
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.|The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: G
authors.initials: G
authors.lastname: Fung
authors.affiliation: Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.|The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: D J
authors.initials: DJ
authors.lastname: Granville
authors.affiliation: Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.|The Centre for Heart and Lung Innovation, St Paul's Hospital, Vancouver, BC, Canada.
authors.forename: D
authors.initials: D
authors.lastname: Yang
mesh_terms: D000818:Animals; D017209:Apoptosis; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D039603:Eukaryotic Initiation Factor-4G / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D000067556:Internal Ribosome Entry Sites / Q000235:genetics*; D051379:Mice; D008805:Mice, Inbred A; D014176:Protein Biosynthesis; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication
chemical_list: C491300:EIF4G2 protein, human; D039603:Eukaryotic Initiation Factor-4G; D000067556:Internal Ribosome Entry Sites; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; C057400:picornain 2A, Picornavirus