Use of a small DNA virus model to investigate mechanisms of CpG dinucleotide-induced attenuation of virus replication
| العنوان: | Use of a small DNA virus model to investigate mechanisms of CpG dinucleotide-induced attenuation of virus replication |
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| المؤلفون: | Peter Simmonds, Lisa Loew, Jeremy Ratcliff, Dung Nguyen, Niluka Goonawardane |
| المصدر: | The Journal of General Virology |
| بيانات النشر: | Microbiology Society, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, minute virus of mice, viruses, Antiviral protein, Viral Nonstructural Proteins, Biology, Virus Replication, codon optimization, Viral Proteins, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Virology, Humans, Codon, Gene, Viral Structural Proteins, Base Composition, Animal, parvovirus, RNA-Binding Proteins, RNA, DNA virus, CpG dinucleotide, biology.organism_classification, TpA dinucleotide, 030104 developmental biology, chemistry, Viral replication, CpG site, A549 Cells, zinc antiviral protein, Small DNA Viruses, Mutation, RNA, Viral, Dinucleoside Phosphates, 030217 neurology & neurosurgery, DNA, Minute virus of mice, Research Article |
| الوصف: | Suppression of the CpG dinucleotide is widespread in RNA viruses infecting vertebrates and plants, and in the genomes of retroviruses and small mammalian DNA viruses. The functional basis for CpG suppression in the latter was investigated through the construction of mutants of the parvovirus, minute virus of mice (MVM) with increased CpG or TpA dinucleotides in the VP gene. CpG-high mutants displayed extraordinary attenuation in A9 cells compared to wild-type MVM (>six logs), while TpA elevation showed no replication effect. Attenuation was independent of Toll-like receptor 9 and STING-mediated DNA recognition pathways and unrelated to effects on translation efficiency. While translation from codon-optimized VP RNA was enhanced in a cell-free assay, MVM containing this sequence was highly attenuated. Further mutational analysis indicated that this arose through its increased numbers of CpG dinucleotides (7→70) and separately from its increased G+C content (42.3→57.4 %), which independently attenuated replication. CpG-high viruses showed impaired NS mRNA expression by qPCR and reduced NS and particularly VP protein expression detected by immunofluorescence and replication in A549 cells, effects reversed in zinc antiviral protein (ZAP) knockout cells, even though nuclear relocalization of VP remained defective. The demonstrated functional basis for CpG suppression in MVM and potentially other small DNA viruses and the observed intolerance of CpGs in coding sequences, even after codon optimization, has implications for the use of small DNA virus vectors in gene therapy and immunization. |
| تدمد: | 1465-2099 0022-1317 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::22034e624b2314db3b5372739b35d3cc https://doi.org/10.1099/jgv.0.001477 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....22034e624b2314db3b5372739b35d3cc |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14652099 00221317 |
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