Second-site mutations in Borna disease virus overexpressing viral accessory protein X

Poenisch, Marion; Wille, Sandra; Schneider, Urs; Staeheli, Peter

doi:10.1099/vir.0.011841-0

Cited by 3 publications

(3 citation statements)

References 10 publications

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“…Structure modelling ( Supplementary Fig. 2) predicts a highly disordered protein, except for a short N-terminal aliphatic alpha helix, that carries both nuclear and mitochondrial localization domains 27 . Thus, we synthesized three peptides covering the entire amino-acid sequence of the X protein to test each of them separately for neuroprotection (Fig.…”

Section: Articlementioning

confidence: 99%

A viral peptide that targets mitochondria protects against neuronal degeneration in models of Parkinson’s disease

et al. 2014

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Mitochondrial dysfunction is a common feature of many neurodegenerative disorders, notably Parkinson's disease. Consequently, agents that protect mitochondria have strong therapeutic potential. Here, we sought to divert the natural strategy used by Borna disease virus (BDV) to replicate in neurons without causing cell death. We show that the BDV X protein has strong axoprotective properties, thereby protecting neurons from degeneration both in tissue culture and in an animal model of Parkinson's disease, even when expressed alone outside of the viral context. We also show that intranasal administration of a cellpermeable peptide derived from the X protein is neuroprotective. We establish that both the X protein and the X-derived peptide act by buffering mitochondrial damage and inducing enhanced mitochondrial filamentation. Our results open the way to novel therapies for neurodegenerative diseases by targeting mitochondrial dynamics and thus preventing the earliest steps of neurodegenerative processes in axons.

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Section: Articlementioning

confidence: 99%

A viral peptide that targets mitochondria protects against neuronal degeneration in models of Parkinson’s disease

et al. 2014

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“…These findings clearly suggest that downregulation of GFP expression occurred at the level of transcription. Insertion of additional A nucleotides into the UA 7 termination motif might interfere with efficient recognition of the termination signal as previously postulated (10). Northern blot analysis of GFP expression 10 weeks p.i.…”

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confidence: 99%

“…Only a minor population of cell somata showed a clear costaining for GFP and nucleoprotein N (yellow cell body and green dendrites), indicating the existence of a population of mBDV-GFP viruses with severely reduced GFP expression capacities. We recently demonstrated that BDV is able to downregulate transcription of an ectopic X gene by modification of the transcription termination/initiation signals directly upstream of the inserted gene (10). To analyze whether similar mechanisms were responsible for the downregulation of GFP expression in mice, we isolated RNA from the brains of mice sacrificed 4 and 10 weeks after infection with mBDV-GFP and amplified by reverse transcription-PCR (RT-PCR) the complete GFP gene, including the upstream sequences that regulate termination and reinitiation of transcription (Fig.…”

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confidence: 99%

Visualizing Viral Dissemination in the Mouse Nervous System, Using a Green Fluorescent Protein-Expressing Borna Disease Virus Vector

Ackermann

Guelzow

Staeheli

et al. 2010

J Virol

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Borna disease virus (BDV) frequently persists in the brain of infected animals.To analyze viral dissemination in the mouse nervous system, we generated a mouse-adapted virus that expresses green fluorescent protein (GFP). This viral vector supported GFP expression for up to 150 days and possessed an extraordinary staining capacity, visualizing complete dendritic arbors as well as individual axonal fibers of infected neurons. GFP-positive cells were first detected in cortical areas from where the virus disseminated through the entire central nervous system (CNS). Late in infection, GFP expression was found in the sciatic nerve, demonstrating viral spread from the central to the peripheral nervous system. Borna disease virus (BDV) is a neurotropic, enveloped virus with a nonsegmented negative-strand RNA genome (2). It naturally infects the central nervous system (CNS) of a broad range of mammalian species (14), where it efficiently establishes persistence in neuronal and nonneuronal cells (4). We recently reported the recovery of a recombinant BDV from cDNA (7, 13) that expresses the green fluorescent protein (GFP) from an additional transcription unit integrated near the 5Ј end of its genome (12). The first-generation GFP vector originating from tissue culture-adapted BDV strain He/80 (13) was severely attenuated in the CNS of adult rats (12) and unable to productively infect mice (our unpublished data). Although mice are resistant to infections with primary BDV isolates, the virus can be adapted to gain replication competence in the CNS of these animals (6, 9). After serial passage of recombinant BDV strain He/80 in the brains of MRL mice, we identified five point mutations that confer replication competence in mice (1). Three of these mutations cause amino acid changes, two in the polymerase L and one in the phosphoprotein P. The other two mutations are silent and located within putative regulatory sequences flanking the initiation codon of the X gene. In a previous study we showed that incorporation of the two adaptive mutations into the L gene (L RD ) improves the growth properties of the GFP-expressing virus in the CNS of rats (12), but the virus remained severely growth retarded in mice (our unpublished data). The mutation in the P gene was previously shown to reduce the sensitivity of the viral polymerase complex for the inhibitory activity of X (1). A recombinant virus carrying this P mutation in combination with the two mutations in L (BDV-P K L RD ) showed strongly enhanced replication speed in mice, whereas the silent mutations within the X gene were found to reduce replication speed and pathogenicity of BDV-P K L RD in MRL mice (1) and rats (our unpublished data). In the present study we incorporated all five adaptive mutations into the BDV genome in order to generate a fully mouse-adapted GFP-expressing BDV vector, designated mBDV-GFP. We found that mBDV-GFP productively infected the central and the peripheral nervous systems of C57BL/6 mice and expressed easily detectable amounts of GFP during the e...

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Two second-site mutations compensate the engineered mutation of R7A in vesicular stomatitis virus nucleocapsid protein

Chen

Zhong

et al. 2016

Virus Research

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Second-site mutations in Borna disease virus overexpressing viral accessory protein X

Cited by 3 publications

References 10 publications

A viral peptide that targets mitochondria protects against neuronal degeneration in models of Parkinson’s disease

A viral peptide that targets mitochondria protects against neuronal degeneration in models of Parkinson’s disease

Visualizing Viral Dissemination in the Mouse Nervous System, Using a Green Fluorescent Protein-Expressing Borna Disease Virus Vector

Two second-site mutations compensate the engineered mutation of R7A in vesicular stomatitis virus nucleocapsid protein

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