Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2
Background/Aims: Epstein-Barr virus (EBV) BFLF2, the homologue of herpes simplex virus 1 (HSV-1) UL31, is crucial for the efficient viral DNA packaging and primary egress across the nuclear membrane. However, we still do not know its subcellular transport mechanisms. Methods: Interspecies heterokary...
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| Format: | Article |
| Language: | English |
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Cell Physiol Biochem Press GmbH & Co KG
2018-11-01
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| Series: | Cellular Physiology and Biochemistry |
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| Online Access: | https://www.karger.com/Article/FullText/495641 |
| _version_ | 1819073630755094528 |
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| author | Meili Li Tao Chen Xingmei Zou Zuo Xu Yuanfang Wang Ping Wang Xiaowen Ou Yiwen Li Daixiong Chen Tao Peng Yao Wang Mingsheng Cai |
| author_facet | Meili Li Tao Chen Xingmei Zou Zuo Xu Yuanfang Wang Ping Wang Xiaowen Ou Yiwen Li Daixiong Chen Tao Peng Yao Wang Mingsheng Cai |
| author_sort | Meili Li |
| collection | DOAJ |
| description | Background/Aims: Epstein-Barr virus (EBV) BFLF2, the homologue of herpes simplex virus 1 (HSV-1) UL31, is crucial for the efficient viral DNA packaging and primary egress across the nuclear membrane. However, we still do not know its subcellular transport mechanisms. Methods: Interspecies heterokaryon assays were utilized to detect the nucleocytoplasmic shuttling of BFLF2, and mutation analysis, plasmid transfection and fluorescence microscopy assays were performed to identify the functional nuclear localization sequence (NLS) and nuclear export sequence (NES) of BFLF2 in live cells. Furthermore, the nuclear import and export of BFLF2 were assessed by confocal microscopy, co-immunoprecipitation and immunoblot assays. Results: BFLF2 was confirmed to shuttle between the nucleus and cytoplasm. Two predicted NESs were shown to be nonfunctional, yet we proved that the nuclear export of BFLF2 was mediated through transporter associated with antigen processing (TAP), but not chromosomal region maintenance 1 (CRM1) dependent pathway. Furthermore, one functional NLS, 22RRLMHPHHRNYTASKASAH40, was identified, and the aa22-23, aa22-25, aa28-30 and aa37-40 had an important role in the nuclear localization of BFLF2. Besides, the nuclear import of BFLF2 was demonstrated through Ran-, importin α7-, importin β1- and transportin-1-dependent mechanism that does not require importin α1, α3 and α5. Conclusion: These works are of significance for the further study of the functions of BFLF2 during EBV infection, as well as for further insights into the design of new antiviral drug target and vaccine development against EBV. |
| first_indexed | 2024-12-21T17:56:41Z |
| format | Article |
| id | doaj.art-ea51ee4877dc40738ddf59687fa1c42f |
| institution | Directory Open Access Journal |
| issn | 1015-8987 1421-9778 |
| language | English |
| last_indexed | 2024-12-21T17:56:41Z |
| publishDate | 2018-11-01 |
| publisher | Cell Physiol Biochem Press GmbH & Co KG |
| record_format | Article |
| series | Cellular Physiology and Biochemistry |
| spelling | doaj.art-ea51ee4877dc40738ddf59687fa1c42f2022-12-21T18:55:11ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782018-11-015141500151710.1159/000495641495641Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2Meili LiTao ChenXingmei ZouZuo XuYuanfang WangPing WangXiaowen OuYiwen LiDaixiong ChenTao PengYao WangMingsheng CaiBackground/Aims: Epstein-Barr virus (EBV) BFLF2, the homologue of herpes simplex virus 1 (HSV-1) UL31, is crucial for the efficient viral DNA packaging and primary egress across the nuclear membrane. However, we still do not know its subcellular transport mechanisms. Methods: Interspecies heterokaryon assays were utilized to detect the nucleocytoplasmic shuttling of BFLF2, and mutation analysis, plasmid transfection and fluorescence microscopy assays were performed to identify the functional nuclear localization sequence (NLS) and nuclear export sequence (NES) of BFLF2 in live cells. Furthermore, the nuclear import and export of BFLF2 were assessed by confocal microscopy, co-immunoprecipitation and immunoblot assays. Results: BFLF2 was confirmed to shuttle between the nucleus and cytoplasm. Two predicted NESs were shown to be nonfunctional, yet we proved that the nuclear export of BFLF2 was mediated through transporter associated with antigen processing (TAP), but not chromosomal region maintenance 1 (CRM1) dependent pathway. Furthermore, one functional NLS, 22RRLMHPHHRNYTASKASAH40, was identified, and the aa22-23, aa22-25, aa28-30 and aa37-40 had an important role in the nuclear localization of BFLF2. Besides, the nuclear import of BFLF2 was demonstrated through Ran-, importin α7-, importin β1- and transportin-1-dependent mechanism that does not require importin α1, α3 and α5. Conclusion: These works are of significance for the further study of the functions of BFLF2 during EBV infection, as well as for further insights into the design of new antiviral drug target and vaccine development against EBV.https://www.karger.com/Article/FullText/495641BFLF2Nuclear transportNuclear localization signal (NLS)Nuclear export signal (NES)ImportinRan-GTP |
| spellingShingle | Meili Li Tao Chen Xingmei Zou Zuo Xu Yuanfang Wang Ping Wang Xiaowen Ou Yiwen Li Daixiong Chen Tao Peng Yao Wang Mingsheng Cai Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2 Cellular Physiology and Biochemistry BFLF2 Nuclear transport Nuclear localization signal (NLS) Nuclear export signal (NES) Importin Ran-GTP |
| title | Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2 |
| title_full | Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2 |
| title_fullStr | Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2 |
| title_full_unstemmed | Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2 |
| title_short | Characterization of the Nucleocytoplasmic Transport Mechanisms of Epstein-Barr Virus BFLF2 |
| title_sort | characterization of the nucleocytoplasmic transport mechanisms of epstein barr virus bflf2 |
| topic | BFLF2 Nuclear transport Nuclear localization signal (NLS) Nuclear export signal (NES) Importin Ran-GTP |
| url | https://www.karger.com/Article/FullText/495641 |
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