Screening of the HBx transactivation domain interacting proteins and the function of interactor Pin1 in HBV replication
Abstract Hepatitis B virus (HBV) X protein (HBx) has been determined to play a crucial role in the replication and transcription of HBV, and its biological functions mainly depend on the interaction with other host proteins. This study aims at screening the proteins that bind to the key functional d...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-93584-z |
| _version_ | 1818999114396860416 |
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| author | Qiaoxia Zhou Libo Yan Baofu Xu Xue’er Wang Xuehong Sun Ning Han Hong Tang Feijun Huang |
| author_facet | Qiaoxia Zhou Libo Yan Baofu Xu Xue’er Wang Xuehong Sun Ning Han Hong Tang Feijun Huang |
| author_sort | Qiaoxia Zhou |
| collection | DOAJ |
| description | Abstract Hepatitis B virus (HBV) X protein (HBx) has been determined to play a crucial role in the replication and transcription of HBV, and its biological functions mainly depend on the interaction with other host proteins. This study aims at screening the proteins that bind to the key functional domain of HBx by integrated proteomics. Proteins that specifically bind to the transactivation domain of HBx were selected by comparing interactors of full-length HBx and HBx-D5 truncation determined by glutathione-S-transferase (GST) pull-down assay combined with mass spectrometry (MS). The function of HBx interactor Pin1 in HBV replication was further investigated by in vitro experiments. In this study, a total of 189 proteins were identified from HepG2 cells that specifically bind to the transactivation domain of HBx by GST pull-down and subsequent MS. After gene ontology (GO) analysis, Pin1 was selected as the protein with the highest score in the largest cluster functioning in protein binding, and also classified into the cluster of proteins with the function of structural molecule activity, which is of great potential to be involved in HBV life cycle. The interaction between Pin1 and HBx has been further confirmed by Ni2+-NTA pulldown assay, co-immunoprecipitation, and immunofluorescence microscopy. HBsAg and HBeAg levels significantly decreased in Pin1 expression inhibited HepG2.2.15 cells. Besides, the inhibition of Pin1 expression in HepG2 cells impeded the restored replication of HBx-deficient HBV repaired by ectopic HBx expression. In conclusion, our study identified Pin1 as an interactor binds to the transactivation domain of HBx, and suggested the potential association between Pin1 and the function of HBx in HBV replication. |
| first_indexed | 2024-12-20T22:12:16Z |
| format | Article |
| id | doaj.art-72c24939c057419794e5340abc3cd0be |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-20T22:12:16Z |
| publishDate | 2021-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-72c24939c057419794e5340abc3cd0be2022-12-21T19:25:08ZengNature PortfolioScientific Reports2045-23222021-07-0111111210.1038/s41598-021-93584-zScreening of the HBx transactivation domain interacting proteins and the function of interactor Pin1 in HBV replicationQiaoxia Zhou0Libo Yan1Baofu Xu2Xue’er Wang3Xuehong Sun4Ning Han5Hong Tang6Feijun Huang7Department of Forensic Pathology, West China School of Preclinical and Forensic Medicine, Sichuan UniversityCenter of Infectious Diseases, West China Hospital of Sichuan UniversityDepartment of Forensic Pathology, West China School of Preclinical and Forensic Medicine, Sichuan UniversityDepartment of Forensic Pathology, West China School of Preclinical and Forensic Medicine, Sichuan UniversityDepartment of Forensic Pathology, West China School of Preclinical and Forensic Medicine, Sichuan UniversityCenter of Infectious Diseases, West China Hospital of Sichuan UniversityCenter of Infectious Diseases, West China Hospital of Sichuan UniversityDepartment of Forensic Pathology, West China School of Preclinical and Forensic Medicine, Sichuan UniversityAbstract Hepatitis B virus (HBV) X protein (HBx) has been determined to play a crucial role in the replication and transcription of HBV, and its biological functions mainly depend on the interaction with other host proteins. This study aims at screening the proteins that bind to the key functional domain of HBx by integrated proteomics. Proteins that specifically bind to the transactivation domain of HBx were selected by comparing interactors of full-length HBx and HBx-D5 truncation determined by glutathione-S-transferase (GST) pull-down assay combined with mass spectrometry (MS). The function of HBx interactor Pin1 in HBV replication was further investigated by in vitro experiments. In this study, a total of 189 proteins were identified from HepG2 cells that specifically bind to the transactivation domain of HBx by GST pull-down and subsequent MS. After gene ontology (GO) analysis, Pin1 was selected as the protein with the highest score in the largest cluster functioning in protein binding, and also classified into the cluster of proteins with the function of structural molecule activity, which is of great potential to be involved in HBV life cycle. The interaction between Pin1 and HBx has been further confirmed by Ni2+-NTA pulldown assay, co-immunoprecipitation, and immunofluorescence microscopy. HBsAg and HBeAg levels significantly decreased in Pin1 expression inhibited HepG2.2.15 cells. Besides, the inhibition of Pin1 expression in HepG2 cells impeded the restored replication of HBx-deficient HBV repaired by ectopic HBx expression. In conclusion, our study identified Pin1 as an interactor binds to the transactivation domain of HBx, and suggested the potential association between Pin1 and the function of HBx in HBV replication.https://doi.org/10.1038/s41598-021-93584-z |
| spellingShingle | Qiaoxia Zhou Libo Yan Baofu Xu Xue’er Wang Xuehong Sun Ning Han Hong Tang Feijun Huang Screening of the HBx transactivation domain interacting proteins and the function of interactor Pin1 in HBV replication Scientific Reports |
| title | Screening of the HBx transactivation domain interacting proteins and the function of interactor Pin1 in HBV replication |
| title_full | Screening of the HBx transactivation domain interacting proteins and the function of interactor Pin1 in HBV replication |
| title_fullStr | Screening of the HBx transactivation domain interacting proteins and the function of interactor Pin1 in HBV replication |
| title_full_unstemmed | Screening of the HBx transactivation domain interacting proteins and the function of interactor Pin1 in HBV replication |
| title_short | Screening of the HBx transactivation domain interacting proteins and the function of interactor Pin1 in HBV replication |
| title_sort | screening of the hbx transactivation domain interacting proteins and the function of interactor pin1 in hbv replication |
| url | https://doi.org/10.1038/s41598-021-93584-z |
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