A Subgenomic Segment of Theiler's Murine Encephalomyelitis Virus RNA Causes Demyelination

Baida, Gleb; Popko, Brian; Wollmann, Robert L.; Stavrou, Spyridon; Lin, Wensheng; Tretiakova, Maria; Krausz, Thomas

doi:10.1128/jvi.02432-07

Cited by 6 publications

(11 citation statements)

References 33 publications

(28 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the EMCV L protein, the zinc finger and the acidic domain are conserved but the C-terminal region encompassing the Ser/Thr-rich domain is lacking. In spite of this difference, L proteins of cardioviruses share the abilities to antagonize IFN production, to affect nucleocytoplasmic trafficking of mRNA and proteins, and to promote nucleoporin hyperphosphorylation (3,16,21,(25)(26)(27)36).Likely as a consequence of mRNA nuclear export inhibition, the TMEV L protein mediates shutoff of host protein synthesis and is very toxic when expressed in cells (2,11,28). In this work, we took advantage of this toxicity to select L mutants that lost the ability to shut off host protein synthesis in order to identify critical domains of the L protein and to test whether the multiple activities of the L protein can be uncoupled.…”

mentioning

confidence: 99%

“…Likely as a consequence of mRNA nuclear export inhibition, the TMEV L protein mediates shutoff of host protein synthesis and is very toxic when expressed in cells (2,11,28). In this work, we took advantage of this toxicity to select L mutants that lost the ability to shut off host protein synthesis in order to identify critical domains of the L protein and to test whether the multiple activities of the L protein can be uncoupled.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Random Mutagenesis Defines a Domain of Theiler's Virus Leader Protein That Is Essential for Antagonism of Nucleocytoplasmic Trafficking and Cytokine Gene Expression

Ricour

Borghese

Sorgeloos

et al. 2009

J Virol

View full text Add to dashboard Cite

The leader protein of cardioviruses, Theiler's murine encephalomyelitis virus (TMEV) and encephalomyocarditis virus (EMCV), is a multifunctional protein known to antagonize type I interferon expression and to interfere with nucleocytoplasmic trafficking of host proteins and mRNA. This protein plays an important role in the capacity of TMEV to establish persistent infection of the central nervous system. Mutant forms of the TMEV leader protein were generated by random mutagenesis and selected after retroviral transduction on the basis of the loss of the highly toxic nature of this protein. Theiler's murine encephalomyelitis virus (TMEV) is a neurotropic picornavirus that belongs to the Cardiovirus genus (reviewed by Brahic et al. [5]). The leader (L) protein of TMEV is a short (76 amino acids), very acidic protein. This protein helps the establishment of persistent TMEV infections in the central nervous system by antagonizing innate host defenses. It inhibits the transcription of type I interferon (IFN) and selected cytokine and chemokine genes, likely through inhibition of IRF-3 dimerization (19,25,28,33,34). It also interferes with nucleocytoplasmic trafficking of cellular proteins and blocks mRNA export from the nucleus (11,28). These activities correlate with the phosphorylation of nucleoporin 98 (Nup98) (28).The sequence of the L protein contains three domains: a zinc finger domain that was shown to bind divalent cations (7), an acidic central domain, and a Ser/Thr-rich domain (see Fig. 2). The L protein encoded by encephalomyocarditis virus (EMCV) shows 35% amino acid identity with the TMEV L protein. In the EMCV L protein, the zinc finger and the acidic domain are conserved but the C-terminal region encompassing the Ser/Thr-rich domain is lacking. In spite of this difference, L proteins of cardioviruses share the abilities to antagonize IFN production, to affect nucleocytoplasmic trafficking of mRNA and proteins, and to promote nucleoporin hyperphosphorylation (3,16,21,(25)(26)(27)36).Likely as a consequence of mRNA nuclear export inhibition, the TMEV L protein mediates shutoff of host protein synthesis and is very toxic when expressed in cells (2,11,28). In this work, we took advantage of this toxicity to select L mutants that lost the ability to shut off host protein synthesis in order to identify critical domains of the L protein and to test whether the multiple activities of the L protein can be uncoupled. MATERIALS AND METHODS Cells and viruses.BHK-21 cells were cultured as previously described (34). BALB/3T3, L929, and Phoenix-Eco cells were cultured in Dulbecco's modified Eagle medium (Gibco) supplemented with 10% fetal bovine serum (MP Biologicals), 100 IU of penicillin/ml, and 100 g of streptomycin/ml. Phoenix-Eco cells were kindly provided by G. Nolan via the ATCC (SD-3444).TMEV derivatives were produced by electroporation of BHK-21 cells (23) with genomic RNA transcribed in vitro from plasmids carrying the corresponding cDNAs. Virus DA1 was produced from plasmid pTMDA1 (10,22,23). Virus TM598 i...

show abstract

mentioning

confidence: 99%

mentioning

confidence: 99%

Random Mutagenesis Defines a Domain of Theiler's Virus Leader Protein That Is Essential for Antagonism of Nucleocytoplasmic Trafficking and Cytokine Gene Expression

Ricour

Borghese

Sorgeloos

et al. 2009

J Virol

View full text Add to dashboard Cite

show abstract

“…The PCR product was digested with NheI and cloned into SpeI-digested pBS246␤ loxP ϳ30 nucleotides downstream from the 3Ј-flanking loxP site. (iii) The transcription terminator sequence and DA L-coding region fragment within pBS246␤ loxP were removed from the resulting plasmid with NotI and inserted into a derivative of pCAGGS that contained a NotI site (1). The resultant DAL transgene was linearized, purified by agarose gel electrophoresis, and used for pronuclear injection.…”

Section: Methodsmentioning

confidence: 99%

“…1A) that contains the chicken ␤-actin promoter followed by a floxed transcription terminator, then followed by the DA L-coding region, and finally followed by the globin poly(A) signal, was constructed in the following way. (i) The plasmid pBS246␤ loxP, which contained an efficient transcription terminator surrounded by loxP sites, was engineered as previously described (1). (ii) The DA L-coding region fragment was amplified from a full-length infectious clone of DA, pDAFL3 (28), using the following primers, which contained NheI enzyme cleavage sites as well as an overlapping Kozak sequence: forward, 5Ј-AGCAACCGCTAGCCACCATGGCTTGCAAA CATGGATAC-3Ј; reverse, 5Ј-AGCAACCGCTAGCTTACTGGGGTTCCAT GACAATATCG-3Ј.…”

Section: Methodsmentioning

confidence: 99%

“…To achieve the intended site-specific and inducible expression, we prepared double transgenic mice, called DA/Cre mice, which carried a tamoxifen-inducible Cre recombinase protein under the control of the myelin proteolipid protein (PLP) transcriptional control element along with a subgenomic region of DA virus. The subgenomic region, which was preceded by a floxed transcription stop, included the DA 5Ј untranslated region (5Ј-UTR), the leader (L) protein-coding region, a region that encodes an out-of-frame protein (L*) important for TMEV-IDD (5,34), and P1 (the capsid-coding region) (1). Interestingly, tamoxifen-treated young DA/Cre mice developed an acute progressive fatal paralysis with abnormalities of the oligodendrocytes and Schwann cells and demyelination but without significant lymphocytic infiltration.…”

mentioning

confidence: 99%

See 1 more Smart Citation

The L-Coding Region of the DA Strain of Theiler's Murine Encephalomyelitis Virus Causes Dysfunction and Death of Myelin-Synthesizing Cells

Ghadge

Wollmann

Baida

et al. 2011

J Virol

View full text Add to dashboard Cite

The DA strain and other members of the TO subgroup of Theiler's murine encephalomyelitis virus (TMEV) induce an early transient subclinical neuronal disease followed by a chronic progressive inflammatory demyelination, with persistence of the virus in the central nervous system (CNS) for the life of the mouse. Although TMEV-induced demyelinating disease (TMEV-IDD) is thought to be immune mediated, there is also evidence that supports a role for the virus in directly inducing demyelination. In order to clarify the function of DA virus genes, we generated a transgenic mouse that had tamoxifen-inducible expression of the DA L-coding region in oligodendrocytes (and Schwann cells), a cell type in which the virus is known to persist. Tamoxifen Theiler's murine encephalomyelitis virus (TMEV) is a member of the Theilovirus species of the Cardiovirus genus of the family of Picornaviridae (reviewed in reference 20). The Encephalomyocarditis virus species of the Cardiovirus genus includes the closely related encephalomyocarditis virus (EMCV) and mengovirus (MV). TMEV strains can be divided into two subgroups on the basis of their differing biologic properties. The GDVII strain and other members of the GDVII subgroup are highly virulent and produce a rapidly fatal neuronal infection in mice, with no persistence of the virus. In contrast, DA, BeAn, and other members of the less virulent TO subgroup induce an early transient subclinical neuronal disease followed by a chronic progressive inflammatory demyelination, with persistence of the virus in the central nervous system (CNS) for the life of the mouse. During TMEV-induced demyelinating disease (TMEV-IDD), relatively large amounts of the TMEV genome can be detected in oligodendrocytes and microglia, with little evidence of viral antigen or infectious virus, i.e., there is a restricted expression of DA viral proteins. TMEV-IDD is an excellent model of multiple sclerosis (MS) because of the similarity in the demyelinating pathologies and because the immune system appears to contribute to pathology in both disorders. Although TMEV-IDD is thought to be immune mediated, there is also evidence that supports a key role for the virus infection in demyelination: the virus persists throughout the demyelinating disease, primarily in oligodendrocytes and microglia; DA virus antigen and virions have been identified in oligodendrocytes (the CNS cells that make myelin), which have a "dying back" pathology (25, 26); and DA virus is known to cause demyelination in infected nude mice (29,30).In order to clarify the functions of selected DA virus genes, we previously generated a mouse that expresses a subgenomic region of the virus as a transgene in myelin-synthesizing cellsoligodendrocytes and Schwann cells. We hypothesized that this transgenic mouse might show a phenotype of interest, thereby clarifying the functions of one or more of the expressed viral genes within myelinating cells. In order to control the time of transgene expression and to avoid tolerance in future immunological studie...

show abstract

Inhibition of mRNA export and dimerization of interferon regulatory factor 3 by Theiler's virus leader protein

Ricour

Delhaye

Hato

et al. 2009

Journal of General Virology

View full text Add to dashboard Cite

Theiler's murine encephalomyelitis virus (TMEV or Theiler's virus) is a neurotropic picornavirus that can persist lifelong in the central nervous system of infected mice, causing a chronic inflammatory demyelinating disease. The leader (L) protein of the virus is an important determinant of viral persistence and has been shown to inhibit transcription of type I interferon (IFN) genes and to cause nucleocytoplasmic redistribution of host proteins. In this study, it was shown that expression of the L protein shuts off synthesis of the reporter proteins green fluorescent protein and firefly luciferase, suggesting that it induces a global shut-off of host protein expression. The L protein did not inhibit transcription or translation of the reporter genes, but blocked cellular mRNA export from the nucleus. This activity correlated with the phosphorylation of nucleoporin 98 (Nup98), an essential component of the nuclear pore complex. In contrast, the data confirmed that the L protein inhibited IFN expression at the transcriptional level, and showed that transcription of other chemokine or cytokine genes was affected by the L protein. This transcriptional inhibition correlated with inhibition of interferon regulatory factor 3 (IRF-3) dimerization. Whether inhibition of IRF-3 dimerization and dysfunction of the nuclear pore complex are related phenomena remains an open question. In vivo, IFN antagonism appears to be an important role of the L protein early in infection, as a virus bearing a mutation in the zinc finger of the L protein replicated as efficiently as the wild-type virus in type I IFN receptor-deficient mice, but had impaired fitness in IFNcompetent mice.

show abstract

A Subgenomic Segment of Theiler's Murine Encephalomyelitis Virus RNA Causes Demyelination

Cited by 6 publications

References 33 publications

Random Mutagenesis Defines a Domain of Theiler's Virus Leader Protein That Is Essential for Antagonism of Nucleocytoplasmic Trafficking and Cytokine Gene Expression

Random Mutagenesis Defines a Domain of Theiler's Virus Leader Protein That Is Essential for Antagonism of Nucleocytoplasmic Trafficking and Cytokine Gene Expression

The L-Coding Region of the DA Strain of Theiler's Murine Encephalomyelitis Virus Causes Dysfunction and Death of Myelin-Synthesizing Cells

Inhibition of mRNA export and dimerization of interferon regulatory factor 3 by Theiler's virus leader protein

Contact Info

Product

Resources

About