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Determination of a phosphorylation site in Nipah virus nucleoprotein and its involvement in virus transcription
Many viruses use their host’s cellular machinery to regulate the functions of viral proteins. The phosphorylation of viral proteins is known to play a role in genome transcription and replication in paramyxoviruses. The paramyxovirus nucleoprotein (N), the most abundant protein in infected cells, is...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Society for General Microbiology
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3353384/ https://www.ncbi.nlm.nih.gov/pubmed/21613447 http://dx.doi.org/10.1099/vir.0.032342-0 |
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author | Huang, Mingshu Sato, Hiroki Hagiwara, Kyoji Watanabe, Akira Sugai, Akihiro Ikeda, Fusako Kozuka-Hata, Hiroko Oyama, Masaaki Yoneda, Misako Kai, Chieko |
author_facet | Huang, Mingshu Sato, Hiroki Hagiwara, Kyoji Watanabe, Akira Sugai, Akihiro Ikeda, Fusako Kozuka-Hata, Hiroko Oyama, Masaaki Yoneda, Misako Kai, Chieko |
author_sort | Huang, Mingshu |
collection | PubMed |
description | Many viruses use their host’s cellular machinery to regulate the functions of viral proteins. The phosphorylation of viral proteins is known to play a role in genome transcription and replication in paramyxoviruses. The paramyxovirus nucleoprotein (N), the most abundant protein in infected cells, is a component of the N–RNA complex and supports the transcription and replication of virus mRNA and genomic RNA. Recently, we reported that the phosphorylation of measles virus N is involved in the regulation of viral RNA synthesis. In this study, we report a rapid turnover of phosphorylation in the Nipah virus N (NiV-N). The phosphorylated NiV-N was hardly detectable in steady-state cells, but was detected after inhibition of cellular protein phosphatases. We identified a phosphorylated serine residue at Ser451 of NiV-N by peptide mass fingerprinting by electrospray ionization–quadrupole time-of-flight mass spectrometry. In the NiV minigenome assay, using luciferase as a reporter gene, the substitution of Ser451 for alanine in NiV-N resulted in a reduction in luciferase activity of approximately 45 % compared with the wild-type protein. Furthermore, the substitution of Ser451 for glutamic acid, which mimics a phosphoserine, led to a more significant decrease in luciferase activity – approximately 81 %. Northern blot analysis showed that both virus transcription and replication were reduced by these mutations. These results suggest that a rapid turnover of the phosphorylation of NiV-N plays an important role in virus transcription and replication. |
format | Online Article Text |
id | pubmed-3353384 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Society for General Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-33533842012-05-29 Determination of a phosphorylation site in Nipah virus nucleoprotein and its involvement in virus transcription Huang, Mingshu Sato, Hiroki Hagiwara, Kyoji Watanabe, Akira Sugai, Akihiro Ikeda, Fusako Kozuka-Hata, Hiroko Oyama, Masaaki Yoneda, Misako Kai, Chieko J Gen Virol Animal Many viruses use their host’s cellular machinery to regulate the functions of viral proteins. The phosphorylation of viral proteins is known to play a role in genome transcription and replication in paramyxoviruses. The paramyxovirus nucleoprotein (N), the most abundant protein in infected cells, is a component of the N–RNA complex and supports the transcription and replication of virus mRNA and genomic RNA. Recently, we reported that the phosphorylation of measles virus N is involved in the regulation of viral RNA synthesis. In this study, we report a rapid turnover of phosphorylation in the Nipah virus N (NiV-N). The phosphorylated NiV-N was hardly detectable in steady-state cells, but was detected after inhibition of cellular protein phosphatases. We identified a phosphorylated serine residue at Ser451 of NiV-N by peptide mass fingerprinting by electrospray ionization–quadrupole time-of-flight mass spectrometry. In the NiV minigenome assay, using luciferase as a reporter gene, the substitution of Ser451 for alanine in NiV-N resulted in a reduction in luciferase activity of approximately 45 % compared with the wild-type protein. Furthermore, the substitution of Ser451 for glutamic acid, which mimics a phosphoserine, led to a more significant decrease in luciferase activity – approximately 81 %. Northern blot analysis showed that both virus transcription and replication were reduced by these mutations. These results suggest that a rapid turnover of the phosphorylation of NiV-N plays an important role in virus transcription and replication. Society for General Microbiology 2011-09 /pmc/articles/PMC3353384/ /pubmed/21613447 http://dx.doi.org/10.1099/vir.0.032342-0 Text en © 2011 SGM http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Animal Huang, Mingshu Sato, Hiroki Hagiwara, Kyoji Watanabe, Akira Sugai, Akihiro Ikeda, Fusako Kozuka-Hata, Hiroko Oyama, Masaaki Yoneda, Misako Kai, Chieko Determination of a phosphorylation site in Nipah virus nucleoprotein and its involvement in virus transcription |
title | Determination of a phosphorylation site in Nipah virus nucleoprotein and its involvement in virus transcription |
title_full | Determination of a phosphorylation site in Nipah virus nucleoprotein and its involvement in virus transcription |
title_fullStr | Determination of a phosphorylation site in Nipah virus nucleoprotein and its involvement in virus transcription |
title_full_unstemmed | Determination of a phosphorylation site in Nipah virus nucleoprotein and its involvement in virus transcription |
title_short | Determination of a phosphorylation site in Nipah virus nucleoprotein and its involvement in virus transcription |
title_sort | determination of a phosphorylation site in nipah virus nucleoprotein and its involvement in virus transcription |
topic | Animal |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3353384/ https://www.ncbi.nlm.nih.gov/pubmed/21613447 http://dx.doi.org/10.1099/vir.0.032342-0 |
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