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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Autores principales:	Qiaoxia Zhou, Libo Yan, Baofu Xu, Xue’er Wang, Xuehong Sun, Ning Han, Hong Tang, Feijun Huang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/72c24939c057419794e5340abc3cd0be
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spelling	oai:doaj.org-article:72c24939c057419794e5340abc3cd0be2021-12-02T16:14:55ZScreening of the HBx transactivation domain interacting proteins and the function of interactor Pin1 in HBV replication10.1038/s41598-021-93584-z2045-2322https://doaj.org/article/72c24939c057419794e5340abc3cd0be2021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-93584-zhttps://doaj.org/toc/2045-2322Abstract 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.Qiaoxia ZhouLibo YanBaofu XuXue’er WangXuehong SunNing HanHong TangFeijun HuangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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
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.
format	article
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
title	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_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_sort	screening of the hbx transactivation domain interacting proteins and the function of interactor pin1 in hbv replication
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/72c24939c057419794e5340abc3cd0be
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Screening of the HBx transactivation domain interacting proteins and the function of interactor Pin1 in HBV replication

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