Analysis of the Physicochemical Properties, Replication and Pathophysiology of a Massively Glycosylated Hepatitis B Virus HBsAg Escape Mutant

Mutations in HBsAg, the surface antigen of the hepatitis B virus (HBV), might affect the serum HBV DNA level of HBV-infected patients, since the reverse transcriptase (RT) domain of HBV polymerase overlaps with the HBsAg-coding region. We previously identified a diagnostic escape mutant (W3S) HBV th...

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Autores principales:	Md. Golzar Hossain, Yadarat Suwanmanee, Kaili Du, Keiji Ueda
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	hepatitis B virus replication HBsAg diagnostic escape mutants glycosylation egress Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/f2ee044c88f74e589b2f28b1c267f6c4
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spelling	oai:doaj.org-article:f2ee044c88f74e589b2f28b1c267f6c42021-11-25T19:14:37ZAnalysis of the Physicochemical Properties, Replication and Pathophysiology of a Massively Glycosylated Hepatitis B Virus HBsAg Escape Mutant10.3390/v131123281999-4915https://doaj.org/article/f2ee044c88f74e589b2f28b1c267f6c42021-11-01T00:00:00Zhttps://www.mdpi.com/1999-4915/13/11/2328https://doaj.org/toc/1999-4915Mutations in HBsAg, the surface antigen of the hepatitis B virus (HBV), might affect the serum HBV DNA level of HBV-infected patients, since the reverse transcriptase (RT) domain of HBV polymerase overlaps with the HBsAg-coding region. We previously identified a diagnostic escape mutant (W3S) HBV that produces massively glycosylated HBsAg. In this study, we constructed an HBV-producing vector that expresses W3S HBs (pHB-W3S) along with a wild-type HBV-producing plasmid (pHB-WT) in order to analyze the physicochemical properties, replication, and antiviral drug response of the mutant. Transfection of either pHB-WT or W3S into HepG2 cells yielded similar CsCl density profiles and eAg expression, as did transfection of a glycosylation defective mutant, pHB-W3S (N146G), in which a glycosylation site at the 146aa asparagine (N) site of HBs was mutated to glycine (G). Virion secretion, however, seemed to be severely impaired in cases of pHB-W3S and pHB-W3S (N146G), compared with pHB-WT, as determined by qPCR and Southern blot analysis. Furthermore, inhibition of glycosylation using tunicamycin<sup>TM</sup> on wild-type HBV production also reduced the virion secretion. These results suggested that the HBV core and Dane particle could be formed either by massively glycosylated or glycosylation-defective HBsAg, but reduced and/or almost completely blocked the virion secretion efficiency, indicating that balanced glycosylation of HBsAg is required for efficient release of HBV, and mutations inducing an imbalanced glycosylation of HBs would cause the virion to become stuck in the cells, which might be associated with various pathogeneses due to HBV infection.Md. Golzar HossainYadarat SuwanmaneeKaili DuKeiji UedaMDPI AGarticlehepatitis B virusreplicationHBsAg diagnostic escape mutantsglycosylationegressMicrobiologyQR1-502ENViruses, Vol 13, Iss 2328, p 2328 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	hepatitis B virus replication HBsAg diagnostic escape mutants glycosylation egress Microbiology QR1-502
spellingShingle	hepatitis B virus replication HBsAg diagnostic escape mutants glycosylation egress Microbiology QR1-502 Md. Golzar Hossain Yadarat Suwanmanee Kaili Du Keiji Ueda Analysis of the Physicochemical Properties, Replication and Pathophysiology of a Massively Glycosylated Hepatitis B Virus HBsAg Escape Mutant
description	Mutations in HBsAg, the surface antigen of the hepatitis B virus (HBV), might affect the serum HBV DNA level of HBV-infected patients, since the reverse transcriptase (RT) domain of HBV polymerase overlaps with the HBsAg-coding region. We previously identified a diagnostic escape mutant (W3S) HBV that produces massively glycosylated HBsAg. In this study, we constructed an HBV-producing vector that expresses W3S HBs (pHB-W3S) along with a wild-type HBV-producing plasmid (pHB-WT) in order to analyze the physicochemical properties, replication, and antiviral drug response of the mutant. Transfection of either pHB-WT or W3S into HepG2 cells yielded similar CsCl density profiles and eAg expression, as did transfection of a glycosylation defective mutant, pHB-W3S (N146G), in which a glycosylation site at the 146aa asparagine (N) site of HBs was mutated to glycine (G). Virion secretion, however, seemed to be severely impaired in cases of pHB-W3S and pHB-W3S (N146G), compared with pHB-WT, as determined by qPCR and Southern blot analysis. Furthermore, inhibition of glycosylation using tunicamycin<sup>TM</sup> on wild-type HBV production also reduced the virion secretion. These results suggested that the HBV core and Dane particle could be formed either by massively glycosylated or glycosylation-defective HBsAg, but reduced and/or almost completely blocked the virion secretion efficiency, indicating that balanced glycosylation of HBsAg is required for efficient release of HBV, and mutations inducing an imbalanced glycosylation of HBs would cause the virion to become stuck in the cells, which might be associated with various pathogeneses due to HBV infection.
format	article
author	Md. Golzar Hossain Yadarat Suwanmanee Kaili Du Keiji Ueda
author_facet	Md. Golzar Hossain Yadarat Suwanmanee Kaili Du Keiji Ueda
author_sort	Md. Golzar Hossain
title	Analysis of the Physicochemical Properties, Replication and Pathophysiology of a Massively Glycosylated Hepatitis B Virus HBsAg Escape Mutant
title_short	Analysis of the Physicochemical Properties, Replication and Pathophysiology of a Massively Glycosylated Hepatitis B Virus HBsAg Escape Mutant
title_full	Analysis of the Physicochemical Properties, Replication and Pathophysiology of a Massively Glycosylated Hepatitis B Virus HBsAg Escape Mutant
title_fullStr	Analysis of the Physicochemical Properties, Replication and Pathophysiology of a Massively Glycosylated Hepatitis B Virus HBsAg Escape Mutant
title_full_unstemmed	Analysis of the Physicochemical Properties, Replication and Pathophysiology of a Massively Glycosylated Hepatitis B Virus HBsAg Escape Mutant
title_sort	analysis of the physicochemical properties, replication and pathophysiology of a massively glycosylated hepatitis b virus hbsag escape mutant
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/f2ee044c88f74e589b2f28b1c267f6c4
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Analysis of the Physicochemical Properties, Replication and Pathophysiology of a Massively Glycosylated Hepatitis B Virus HBsAg Escape Mutant

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