Encephalomyocarditis Virus 2A Protein Inhibited Apoptosis by Interaction with Annexin A2 through JNK/c-Jun Pathway

Abstract

Encephalomyocarditis virus can cause myocarditis and encephalitis in pigs and other mammals, thus posing a potential threat to public health safety. The 2A protein is an important virulence factor of EMCV. Previous studies have shown that the 2A protein may be related to the inhibition of apoptosis by virus, but its specific molecular mechanism is not clear. In this study, the 2A protein was expressed in Escherichia coli in order to find interacting cell proteins. A pull down assay, coupled with mass spectrometry, revealed that the 2A protein possibly interacted with annexin A2. Co-immunoprecipitation assays and confocal imaging analysis further demonstrated that the 2A protein interacted with annexin A2 in cells. In reducing the expression of annexin A2 by siRNA, the ability of the 2A protein to inhibit apoptosis was weakened and the proliferation of EMCV was slowed down. These results suggest that annexin A2 is closely related to the inhibition of apoptosis by 2A. Furthermore, both RT-PCR and western blot results showed that the 2A protein requires annexin A2 interaction to inhibit apoptosis via JNK/c-Jun pathway. Taken together, our data indicate that the 2A protein inhibits apoptosis by interacting with annexin A2 via the JNK/c-Jun pathway. These findings provide insight into the molecular pathogenesis underlying EMCV infection.

Comprehensive Elucidation of the Role of L and 2A Security Proteins on Cell Death during EMCV Infection

“…Moreover, published data showed that the deletion of 2A results in the apoptosis of EMCV-infected BHK-21 cells [ 30 ]. 2A may inhibit the chemical-induced apoptotic death of PK15 and BHK-21 cells via its ectopic expression [ 31 ]. All the described investigations included the study of one protein in a particular cell culture.…”

“…The mechanism of 2A interference in cell death is still unclear. There is only one experimentally based assumption about mechanism: EMCV 2A plays an antiapoptotic role by interaction with Annexin A2 [ 31 ]. Nevertheless, this proposition is based on a 2A-expressing model, not a viral-infecting model.…”

“…Moreover, similar to L protein, 2A was shown to exhibit anti-apoptotic activity, though a different cell line, BHK-21, was used [ 30 ]. Studies examining the ectopic expression of the 2A protein in pig cells (PK15) have also suggested that the 2A protein may have an anti-apoptotic role by potentially interacting with Annexin-A2 [ 31 ].…”

The V protein in oncolytic Newcastle disease virus promotes HepG2 hepatoma cell proliferation at the single-cell level

“…The effects of the transactivator Tas encoded by foamy viruses and expressed after cell cycle arrest or before apoptosis highlight the different mechanisms through which virus–host interactions inhibit cell proliferation [ 21 ]. Encephalomyocarditis virus 2A protein inhibited apoptosis [ 22 ], and human papilloma virus 16 (HPV16) protein E7 increased PKM2 expression and activated the nonglycolytic function of PKM2 to promote cervical cancer cell proliferation [ 23 ]. However, no direct evidence has shown to date that the V protein encoded by NDV regulates host cell proliferation; however, our present work shows that the V protein promotes cancer cell proliferation.…”

Impact of Annexin A2 on virus life cycles

“…The P2 and P3 precursor proteins are processed to generate the non-structural proteins 2A, 2B, 2C, 3A, 3B (Vpg), 3C protease, and the RNA-dependent RNA polymerase 3D ( Palmenberg et al, 1984 ). Among these viral proteins, AnxA2 is known to interact with the virulence protein 2A, which is required for viral pathogenesis and inhibition of apoptosis through the JNK/c-Jun pathway, promoting EMCV replication during the early stage of infection ( Carocci et al, 2011 ; Han et al, 2022 ). ii) Enterovirus 71 (EV71) .…”