A Conserved Phenylalanine Residue of Autographa Californica Multiple Nucleopolyhedrovirus AC75 Protein Is Required for Occlusion Body Formation
Autographa californica multiple nucleopolyhedrovirus (AcMNPV) orf75 (ac75) is a highly conserved gene that is essential for AcMNPV propagation. However, the key domains or residues of the AC75 protein that play a role in viral propagation have not been identified. In this study, sequence alignment r...
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| Format: | Article |
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Frontiers Media S.A.
2021-04-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2021.663506/full |
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| author | Xingang Chen Xingang Chen Jian Yang Jian Yang Xiaoqin Yang Xiaoqin Yang Chengfeng Lei Xiulian Sun Jia Hu |
| author_facet | Xingang Chen Xingang Chen Jian Yang Jian Yang Xiaoqin Yang Xiaoqin Yang Chengfeng Lei Xiulian Sun Jia Hu |
| author_sort | Xingang Chen |
| collection | DOAJ |
| description | Autographa californica multiple nucleopolyhedrovirus (AcMNPV) orf75 (ac75) is a highly conserved gene that is essential for AcMNPV propagation. However, the key domains or residues of the AC75 protein that play a role in viral propagation have not been identified. In this study, sequence alignment revealed that residues Phe-54 and Gln-81 of AC75 were highly conserved among alphabaculoviruses and betabaculoviurses. Thus, Phe-54 and Gln-81 AC75 mutation bacmids were constructed. We found that Gln-81 was not required for viral propagation, whereas mutating Phe-54 reduced budded virus production by 10-fold and impaired occlusion body formation when compared with that of the wild-type AcMNPV. Electron microscopy observations showed that the Phe-54 mutation affected polyhedrin assembly and also occlusion-derived virus embedding, whereas western blot analysis revealed that mutating Phe-54 reduced the amount of AC75 but did not affect the localization of AC75 in infected cells. A protein stability assay showed that the Phe-54 mutation affected AC75 stability. Taken together, Phe-54 was identified as an important residue of AC75, and ac75 is a pivotal gene in budding virus production and occlusion body formation. |
| first_indexed | 2024-12-16T12:22:21Z |
| format | Article |
| id | doaj.art-571e3851265349eaba552aed06544d86 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-16T12:22:21Z |
| publishDate | 2021-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-571e3851265349eaba552aed06544d862022-12-21T22:31:55ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-04-011210.3389/fmicb.2021.663506663506A Conserved Phenylalanine Residue of Autographa Californica Multiple Nucleopolyhedrovirus AC75 Protein Is Required for Occlusion Body FormationXingang Chen0Xingang Chen1Jian Yang2Jian Yang3Xiaoqin Yang4Xiaoqin Yang5Chengfeng Lei6Xiulian Sun7Jia Hu8Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, ChinaCollege of Life Sciences, University of Chinese Academy of Sciences, Beijing, ChinaWuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, ChinaCollege of Life Sciences, University of Chinese Academy of Sciences, Beijing, ChinaWuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, ChinaCollege of Life Sciences, University of Chinese Academy of Sciences, Beijing, ChinaWuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, ChinaWuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, ChinaWuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, ChinaAutographa californica multiple nucleopolyhedrovirus (AcMNPV) orf75 (ac75) is a highly conserved gene that is essential for AcMNPV propagation. However, the key domains or residues of the AC75 protein that play a role in viral propagation have not been identified. In this study, sequence alignment revealed that residues Phe-54 and Gln-81 of AC75 were highly conserved among alphabaculoviruses and betabaculoviurses. Thus, Phe-54 and Gln-81 AC75 mutation bacmids were constructed. We found that Gln-81 was not required for viral propagation, whereas mutating Phe-54 reduced budded virus production by 10-fold and impaired occlusion body formation when compared with that of the wild-type AcMNPV. Electron microscopy observations showed that the Phe-54 mutation affected polyhedrin assembly and also occlusion-derived virus embedding, whereas western blot analysis revealed that mutating Phe-54 reduced the amount of AC75 but did not affect the localization of AC75 in infected cells. A protein stability assay showed that the Phe-54 mutation affected AC75 stability. Taken together, Phe-54 was identified as an important residue of AC75, and ac75 is a pivotal gene in budding virus production and occlusion body formation.https://www.frontiersin.org/articles/10.3389/fmicb.2021.663506/fullAutographa californica multiple nucleopolyhedrovirusAC75Phe-54budded virusocclusion body |
| spellingShingle | Xingang Chen Xingang Chen Jian Yang Jian Yang Xiaoqin Yang Xiaoqin Yang Chengfeng Lei Xiulian Sun Jia Hu A Conserved Phenylalanine Residue of Autographa Californica Multiple Nucleopolyhedrovirus AC75 Protein Is Required for Occlusion Body Formation Frontiers in Microbiology Autographa californica multiple nucleopolyhedrovirus AC75 Phe-54 budded virus occlusion body |
| title | A Conserved Phenylalanine Residue of Autographa Californica Multiple Nucleopolyhedrovirus AC75 Protein Is Required for Occlusion Body Formation |
| title_full | A Conserved Phenylalanine Residue of Autographa Californica Multiple Nucleopolyhedrovirus AC75 Protein Is Required for Occlusion Body Formation |
| title_fullStr | A Conserved Phenylalanine Residue of Autographa Californica Multiple Nucleopolyhedrovirus AC75 Protein Is Required for Occlusion Body Formation |
| title_full_unstemmed | A Conserved Phenylalanine Residue of Autographa Californica Multiple Nucleopolyhedrovirus AC75 Protein Is Required for Occlusion Body Formation |
| title_short | A Conserved Phenylalanine Residue of Autographa Californica Multiple Nucleopolyhedrovirus AC75 Protein Is Required for Occlusion Body Formation |
| title_sort | conserved phenylalanine residue of autographa californica multiple nucleopolyhedrovirus ac75 protein is required for occlusion body formation |
| topic | Autographa californica multiple nucleopolyhedrovirus AC75 Phe-54 budded virus occlusion body |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2021.663506/full |
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