Mutation Profiles of eGFP-Tagged Small Ruminant Morbillivirus During 45 Serial Passages in Ribavirin-Treated Cells
Small ruminant morbillivirus (SRMV), formerly known as peste-des-petits-ruminants virus, classified into the genus Morbillivirus in the family Paramyxoviridae. Its L protein functions as the RNA-dependent RNA polymerases (RdRp) during viral replication. Due to the absence of efficient proofreading a...
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Frontiers Media S.A.
2021-07-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2021.690204/full |
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| author | Fuxiao Liu Yanli Zou Lin Li Chunju Liu Xiaodong Wu |
| author_facet | Fuxiao Liu Yanli Zou Lin Li Chunju Liu Xiaodong Wu |
| author_sort | Fuxiao Liu |
| collection | DOAJ |
| description | Small ruminant morbillivirus (SRMV), formerly known as peste-des-petits-ruminants virus, classified into the genus Morbillivirus in the family Paramyxoviridae. Its L protein functions as the RNA-dependent RNA polymerases (RdRp) during viral replication. Due to the absence of efficient proofreading activity in their RdRps, various RNA viruses reveal high mutation frequencies, making them evolve rapidly during serial passages in cells, especially treated with a certain mutagen, like ribavirin. We have previously rescued a recombinant enhanced green fluorescence protein-tagged SRMV (rSRMV-eGFP) using reverse genetics. In this study, the rSRMV-eGFP was subjected to serial passages in ribavirin-treated cells. Due to the ribavirin-exerted selective pressure, it was speculated that viral progenies would form quasispecies after dozens of passages. Viral progenies at passage-10, -20, -30, -40, and -50 were separately subjected to next-generation sequencing (NGS), consequently revealing a total of 34 single-nucleotide variations, including five synonymous, 21 missense, and one non-sense mutations. The L sequence was found to harbor eight missense mutations during serial passaging. It was speculated that at least one high-fidelity variant was present in viral quasispecies at passage-50. If purified from the population of viral progenies, this putative variant would contribute to clarifying a molecular mechanism in viral high-fidelity replication in vitro. |
| first_indexed | 2024-12-22T13:02:02Z |
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| institution | Directory Open Access Journal |
| issn | 2297-1769 |
| language | English |
| last_indexed | 2024-12-22T13:02:02Z |
| publishDate | 2021-07-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Veterinary Science |
| spelling | doaj.art-fa83c44873604245bba236d1add1f8d42022-12-21T18:24:59ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692021-07-01810.3389/fvets.2021.690204690204Mutation Profiles of eGFP-Tagged Small Ruminant Morbillivirus During 45 Serial Passages in Ribavirin-Treated CellsFuxiao Liu0Yanli Zou1Lin Li2Chunju Liu3Xiaodong Wu4College of Veterinary Medicine, Qingdao Agricultural University, Qingdao, ChinaOIE Reference Laboratory for Peste des Petits Ruminants, China Animal Health and Epidemiology Center, Qingdao, ChinaOIE Reference Laboratory for Peste des Petits Ruminants, China Animal Health and Epidemiology Center, Qingdao, ChinaOIE Reference Laboratory for Peste des Petits Ruminants, China Animal Health and Epidemiology Center, Qingdao, ChinaOIE Reference Laboratory for Peste des Petits Ruminants, China Animal Health and Epidemiology Center, Qingdao, ChinaSmall ruminant morbillivirus (SRMV), formerly known as peste-des-petits-ruminants virus, classified into the genus Morbillivirus in the family Paramyxoviridae. Its L protein functions as the RNA-dependent RNA polymerases (RdRp) during viral replication. Due to the absence of efficient proofreading activity in their RdRps, various RNA viruses reveal high mutation frequencies, making them evolve rapidly during serial passages in cells, especially treated with a certain mutagen, like ribavirin. We have previously rescued a recombinant enhanced green fluorescence protein-tagged SRMV (rSRMV-eGFP) using reverse genetics. In this study, the rSRMV-eGFP was subjected to serial passages in ribavirin-treated cells. Due to the ribavirin-exerted selective pressure, it was speculated that viral progenies would form quasispecies after dozens of passages. Viral progenies at passage-10, -20, -30, -40, and -50 were separately subjected to next-generation sequencing (NGS), consequently revealing a total of 34 single-nucleotide variations, including five synonymous, 21 missense, and one non-sense mutations. The L sequence was found to harbor eight missense mutations during serial passaging. It was speculated that at least one high-fidelity variant was present in viral quasispecies at passage-50. If purified from the population of viral progenies, this putative variant would contribute to clarifying a molecular mechanism in viral high-fidelity replication in vitro.https://www.frontiersin.org/articles/10.3389/fvets.2021.690204/fullrSRMV-eGFPnext-generation sequencingsingle-nucleotide variationmutationL proteinhigh-fidelity |
| spellingShingle | Fuxiao Liu Yanli Zou Lin Li Chunju Liu Xiaodong Wu Mutation Profiles of eGFP-Tagged Small Ruminant Morbillivirus During 45 Serial Passages in Ribavirin-Treated Cells Frontiers in Veterinary Science rSRMV-eGFP next-generation sequencing single-nucleotide variation mutation L protein high-fidelity |
| title | Mutation Profiles of eGFP-Tagged Small Ruminant Morbillivirus During 45 Serial Passages in Ribavirin-Treated Cells |
| title_full | Mutation Profiles of eGFP-Tagged Small Ruminant Morbillivirus During 45 Serial Passages in Ribavirin-Treated Cells |
| title_fullStr | Mutation Profiles of eGFP-Tagged Small Ruminant Morbillivirus During 45 Serial Passages in Ribavirin-Treated Cells |
| title_full_unstemmed | Mutation Profiles of eGFP-Tagged Small Ruminant Morbillivirus During 45 Serial Passages in Ribavirin-Treated Cells |
| title_short | Mutation Profiles of eGFP-Tagged Small Ruminant Morbillivirus During 45 Serial Passages in Ribavirin-Treated Cells |
| title_sort | mutation profiles of egfp tagged small ruminant morbillivirus during 45 serial passages in ribavirin treated cells |
| topic | rSRMV-eGFP next-generation sequencing single-nucleotide variation mutation L protein high-fidelity |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2021.690204/full |
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