Binding Interactions between the Encephalomyocarditis Virus Leader and Protein 2A

Petty, Ryan V.; Basta, Holly A.; Bacot‐Davis, Valjean R.; Brown, Bradley A.; Palmenberg, Ann C.

doi:10.1128/jvi.02148-14

Cited by 12 publications

(21 citation statements)

References 31 publications

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“…The viral L protein, encoded at the very N-terminus of the polyprotein is acidic and known to functionally interact with the basic 2A protein during virus infection (34). However, whether L has any effect on PRF has not been investigated.…”

Section: Resultsmentioning

confidence: 99%

Characterization of the stimulators of protein-directed ribosomal frameshifting in Theiler's murine encephalomyelitis virus

Napthine

Bell

Hill

et al. 2019

Nucleic Acids Research

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Many viruses utilize programmed –1 ribosomal frameshifting (–1 PRF) to express additional proteins or to produce frameshift and non-frameshift protein products at a fixed stoichiometric ratio. PRF is also utilized in the expression of a small number of cellular genes. Frameshifting is typically stimulated by signals contained within the mRNA: a ‘slippery’ sequence and a 3′-adjacent RNA structure. Recently, we showed that −1 PRF in encephalomyocarditis virus (EMCV) is trans-activated by the viral 2A protein, leading to a temporal change in PRF efficiency from 0% to 70% during virus infection. Here we analyzed PRF in the related Theiler's murine encephalomyelitis virus (TMEV). We show that 2A is also required for PRF in TMEV and can stimulate PRF to levels as high as 58% in rabbit reticulocyte cell-free translations and 81% during virus infection. We also show that TMEV 2A trans-activates PRF on the EMCV signal but not vice versa. We present an extensive mutational analysis of the frameshift stimulators (mRNA signals and 2A protein) analysing activity in in vitro translation, electrophoretic mobility shift and in vitro ribosome pausing assays. We also investigate the PRF mRNA signal with RNA structure probing. Our results substantially extend previous characterization of protein-stimulated PRF.

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Section: Resultsmentioning

confidence: 99%

Characterization of the stimulators of protein-directed ribosomal frameshifting in Theiler's murine encephalomyelitis virus

Napthine

Bell

Hill

et al. 2019

Nucleic Acids Research

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“…Recombinant GST-tagged L X materials readily recapitulate most of the protein phenotypes in cells and extracts and are typically used to explore these mechanisms (4, 12, 17). With both native (HeLa cytosol) and recombinant (rCrm1) sources, reciprocal pulldowns indicated strong, salt-resistant direct interactions between 3 different cardiovirus (GST-)L X sequences and Crm1/CAS (Fig 2, Fig 3).…”

Section: Discussionmentioning

confidence: 99%

Cardiovirus Leader proteins bind exportins: Implications for virus replication and nucleocytoplasmic trafficking inhibition

2016

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Cardiovirus Leader proteins (LX) inhibit cellular nucleocytoplasmic trafficking by directing host kinases to phosphorylate Phe/Gly-containing nuclear pore proteins (Nups). Resolution of the Mengovirus LM structure bound to Ran GTPase, suggested this complex would further recruit specific exportins (karyopherins), which in turn mediate kinase selection. Pull-down experiments and recombinant complex reconstitution now confirm that Crm1 and CAS exportins form stable dimeric complexes with encephalomyocarditis virus LE, and also larger complexes with LE:Ran. shRNA knockdown studies support this idea. Similar activities could be demonstrated for recombinant LS and LT from Theiloviruses. When mutations were introduced to alter the LE zinc finger domain, acidic domain, or dual phosphorylation sites, there was reduced exportin selection. These regions are not involved in Ran interactions, so the Ran and Crm1 binding sites on LE must be non-overlapping. The involvement of exportins in this mechanism is important to viral replication and the observation of trafficking inhibition by LE.

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“…Localization of L to the nucleus depends on expression of 2A, which contains a NLS in its C-terminus ( Groppo et al, 2011 ). Upon nuclear localization, L interacts with Ran with high affinity and 2A is displaced as the binding sites for 2A and Ran partially overlap ( Petty et al, 2014 ). L from EMCV, TMEV, and SAFV induce hyper-phosphorylation of Nups including Nup62 and Nup98 ( Ricour et al, 2009 ; Ciomperlik et al, 2015 ), likely by recruiting and activating a kinase, which may be facilitated by L binding of exportins Crm1 and CAS ( Ciomperlik et al, 2016 ).…”

Section: Inhibition Of Interferon Production By Cardiovirusesmentioning

confidence: 99%

Innate Immune Detection of Cardioviruses and Viral Disruption of Interferon Signaling

2018

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Cardioviruses are members of the Picornaviridae family and infect a variety of mammals, from mice to humans. Replication of cardioviruses produces double stranded RNA that is detected by helicases in the RIG-I-like receptor family and leads to a signaling cascade to produce type I interferon. Like other viruses within Picornaviridae, however, cardioviruses have evolved several mechanisms to inhibit interferon production. In this review, we summarize recent findings that have uncovered several proteins enabling efficient detection of cardiovirus dsRNA and discuss which cell types may be most important for interferon production in vivo. Additionally, we describe how cardiovirus proteins L, 3C and L∗ disrupt interferon production and antagonize the antiviral activity of interferon effector molecules.

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Binding Interactions between the Encephalomyocarditis Virus Leader and Protein 2A

Cited by 12 publications

References 31 publications

Characterization of the stimulators of protein-directed ribosomal frameshifting in Theiler's murine encephalomyelitis virus

Characterization of the stimulators of protein-directed ribosomal frameshifting in Theiler's murine encephalomyelitis virus

Cardiovirus Leader proteins bind exportins: Implications for virus replication and nucleocytoplasmic trafficking inhibition

Innate Immune Detection of Cardioviruses and Viral Disruption of Interferon Signaling

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