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N(6)-Methyladenosine RNA Modification in Host Cells Regulates Peste des Petits Ruminants Virus Replication
N(6)-methyladenosine (m(6)A) modification is a major RNA epigenetic regulatory mechanism. The dynamics of m(6)A levels in viral genomic RNA and their mRNAs have been shown to have either pro- or antiviral functions, and therefore, m(6)A modifications influence virus-host interactions. Currently, no...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10101086/ https://www.ncbi.nlm.nih.gov/pubmed/36786625 http://dx.doi.org/10.1128/spectrum.02666-22 |
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author | Khan, Owais Tanuj, Gunturu Narasimha Choravada, Divyaprakash R. Rajak, Kaushal Kishore Chandra Sekar, S Lingaraju, Madhu Cholenahalli Dhara, Sujoy K. Gupta, Praveen K. Mishra, Bishnu Prasad Dutt, Triveni Gandham, Ravi Kumar Sajjanar, Basavaraj |
author_facet | Khan, Owais Tanuj, Gunturu Narasimha Choravada, Divyaprakash R. Rajak, Kaushal Kishore Chandra Sekar, S Lingaraju, Madhu Cholenahalli Dhara, Sujoy K. Gupta, Praveen K. Mishra, Bishnu Prasad Dutt, Triveni Gandham, Ravi Kumar Sajjanar, Basavaraj |
author_sort | Khan, Owais |
collection | PubMed |
description | N(6)-methyladenosine (m(6)A) modification is a major RNA epigenetic regulatory mechanism. The dynamics of m(6)A levels in viral genomic RNA and their mRNAs have been shown to have either pro- or antiviral functions, and therefore, m(6)A modifications influence virus-host interactions. Currently, no reports are available on the effect of m(6)A modifications in the genome of Peste des petits ruminants virus (PPRV). In the present study, we took PPRV as a model for nonsegmented negative-sense single-stranded RNA viruses and elucidate the role of m(6)A modification on viral replication. We detected m(6)A-modified sites in the mRNA of the virus and host cells, as well as the PPRV RNA genome. Further, it was found that the level of m(6)A modification in host cells alters the viral gene expression. Knockdown of the METTL3 and FTO genes (encoding the m(6)A RNA modification writer and eraser proteins, respectively) results in alterations of the levels of m(6)A RNA modifications in the host cells. Experiments using these genetically modified clones of host cells infected with PPRV revealed that both higher and lower m(6)A RNA modification in the host cells negatively affect PPRV replication. We found that m(6)A-modified viral transcripts had better stability and translation efficiency compared to the unmodified mRNA. Altogether, from these data, we conclude that the m(6)A modification of RNA regulates PPRV replication. These findings contribute toward a way forward for developing novel antiviral strategies against PPRV by modulating the dynamics of host m(6)A RNA modification. IMPORTANCE Peste des petits ruminants virus (PPRV) causes a severe disease in sheep and goats. PPRV infection is a major problem, causing significant economic losses to small ruminant farmers in regions of endemicity. N(6)-methyladenosine (m(6)A) is an important RNA modification involved in various functions, including virus-host interactions. In the present study, we used stable clones of Vero cells, having knocked down the genes encoding proteins involved in dynamic changes of the levels of m(6)A modification. We also used small-molecule compounds that interfere with m(6)A methylation. This resulted in a platform of host cells with various degrees of m(6)A RNA modification. The host cells with these different microenvironments were useful for studying the effect of m(6)A RNA modification on the expression of viral genes and viral replication. The results pinpoint the level of m(6)A modifications that facilitate the maximum replication of PPRV. These findings will be useful in increasing the virus titers in cultured cells needed for the economical development of the vaccine. Furthermore, the findings have guiding significance for the development of novel antiviral strategies for limiting PPRV replication in infected animals. |
format | Online Article Text |
id | pubmed-10101086 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-101010862023-04-14 N(6)-Methyladenosine RNA Modification in Host Cells Regulates Peste des Petits Ruminants Virus Replication Khan, Owais Tanuj, Gunturu Narasimha Choravada, Divyaprakash R. Rajak, Kaushal Kishore Chandra Sekar, S Lingaraju, Madhu Cholenahalli Dhara, Sujoy K. Gupta, Praveen K. Mishra, Bishnu Prasad Dutt, Triveni Gandham, Ravi Kumar Sajjanar, Basavaraj Microbiol Spectr Research Article N(6)-methyladenosine (m(6)A) modification is a major RNA epigenetic regulatory mechanism. The dynamics of m(6)A levels in viral genomic RNA and their mRNAs have been shown to have either pro- or antiviral functions, and therefore, m(6)A modifications influence virus-host interactions. Currently, no reports are available on the effect of m(6)A modifications in the genome of Peste des petits ruminants virus (PPRV). In the present study, we took PPRV as a model for nonsegmented negative-sense single-stranded RNA viruses and elucidate the role of m(6)A modification on viral replication. We detected m(6)A-modified sites in the mRNA of the virus and host cells, as well as the PPRV RNA genome. Further, it was found that the level of m(6)A modification in host cells alters the viral gene expression. Knockdown of the METTL3 and FTO genes (encoding the m(6)A RNA modification writer and eraser proteins, respectively) results in alterations of the levels of m(6)A RNA modifications in the host cells. Experiments using these genetically modified clones of host cells infected with PPRV revealed that both higher and lower m(6)A RNA modification in the host cells negatively affect PPRV replication. We found that m(6)A-modified viral transcripts had better stability and translation efficiency compared to the unmodified mRNA. Altogether, from these data, we conclude that the m(6)A modification of RNA regulates PPRV replication. These findings contribute toward a way forward for developing novel antiviral strategies against PPRV by modulating the dynamics of host m(6)A RNA modification. IMPORTANCE Peste des petits ruminants virus (PPRV) causes a severe disease in sheep and goats. PPRV infection is a major problem, causing significant economic losses to small ruminant farmers in regions of endemicity. N(6)-methyladenosine (m(6)A) is an important RNA modification involved in various functions, including virus-host interactions. In the present study, we used stable clones of Vero cells, having knocked down the genes encoding proteins involved in dynamic changes of the levels of m(6)A modification. We also used small-molecule compounds that interfere with m(6)A methylation. This resulted in a platform of host cells with various degrees of m(6)A RNA modification. The host cells with these different microenvironments were useful for studying the effect of m(6)A RNA modification on the expression of viral genes and viral replication. The results pinpoint the level of m(6)A modifications that facilitate the maximum replication of PPRV. These findings will be useful in increasing the virus titers in cultured cells needed for the economical development of the vaccine. Furthermore, the findings have guiding significance for the development of novel antiviral strategies for limiting PPRV replication in infected animals. American Society for Microbiology 2023-02-14 /pmc/articles/PMC10101086/ /pubmed/36786625 http://dx.doi.org/10.1128/spectrum.02666-22 Text en Copyright © 2023 Khan et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Article Khan, Owais Tanuj, Gunturu Narasimha Choravada, Divyaprakash R. Rajak, Kaushal Kishore Chandra Sekar, S Lingaraju, Madhu Cholenahalli Dhara, Sujoy K. Gupta, Praveen K. Mishra, Bishnu Prasad Dutt, Triveni Gandham, Ravi Kumar Sajjanar, Basavaraj N(6)-Methyladenosine RNA Modification in Host Cells Regulates Peste des Petits Ruminants Virus Replication |
title | N(6)-Methyladenosine RNA Modification in Host Cells Regulates Peste des Petits Ruminants Virus Replication |
title_full | N(6)-Methyladenosine RNA Modification in Host Cells Regulates Peste des Petits Ruminants Virus Replication |
title_fullStr | N(6)-Methyladenosine RNA Modification in Host Cells Regulates Peste des Petits Ruminants Virus Replication |
title_full_unstemmed | N(6)-Methyladenosine RNA Modification in Host Cells Regulates Peste des Petits Ruminants Virus Replication |
title_short | N(6)-Methyladenosine RNA Modification in Host Cells Regulates Peste des Petits Ruminants Virus Replication |
title_sort | n(6)-methyladenosine rna modification in host cells regulates peste des petits ruminants virus replication |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10101086/ https://www.ncbi.nlm.nih.gov/pubmed/36786625 http://dx.doi.org/10.1128/spectrum.02666-22 |
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