Identification and characterization of a G-quadruplex structure in the pre-core promoter region of hepatitis B virus covalently closed circular DNA

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dc.contributor.authorMeier-Stephenson, Vanessa
dc.contributor.authorBadmalia, Maulik D.
dc.contributor.authorMrozowich, Tyler
dc.contributor.authorLau, Keith C. K.
dc.contributor.authorSchultz, Sarah K.
dc.contributor.authorGemmill, Darren L.
dc.contributor.authorOsiowy, Carla
dc.contributor.authorvan Marle, Guido
dc.contributor.authorCoffin, Carla S.
dc.contributor.authorPatel, Trushar R.
dc.date.accessioned2022-10-28T19:50:11Z
dc.date.available2022-10-28T19:50:11Z
dc.date.issued2021
dc.descriptionOpen access article. Creative Commons Attribution 4.0 International License (CC BY 4.0) appliesen_US
dc.description.abstractApproximately 250 million people worldwide are chronically infected with the hepatitis B virus (HBV) and are at increased risk of developing cirrhosis and hepatocellular carcinoma. The HBV genome persists as covalently closed circular DNA (cccDNA), which serves as the template for all HBV mRNA transcripts. Current nucleos(t)ide analogs used to treat HBV do not directly target the HBV cccDNA genome and thus cannot eradicate HBV infection. Here, we report the discovery of a unique G-quadruplex structure in the pre-core promoter region of the HBV genome that is conserved among nearly all genotypes. This region is central to critical steps in the viral life cycle, including the generation of pregenomic RNA, synthesis of core and polymerase proteins, and genome encapsidation; thus, an increased understanding of the HBV pre-core region may lead to the identification of novel anti-HBV cccDNA targets. We utilized biophysical methods (circular dichroism and small-angle X-ray scattering) to characterize the HBV G-quadruplex and the effect of three distinct G to A mutants. We also used microscale thermophoresis to quantify the binding affinity of G-quadruplex and its mutants with a known quadruplex-binding protein (DHX36). To investigate the physiological relevance of HBV G-quadruplex, we employed assays using DHX36 to pull-down cccDNA and compared HBV infection in HepG2 cells transfected with wild-type and mutant HBV plasmids by monitoring the levels of genomic DNA, pregenomic RNA, and antigens. Further evaluation of this critical host-protein interaction site in the HBV cccDNA genome may facilitate the development of novel anti-HBV therapeutics against the resilient cccDNA template.en_US
dc.identifier.citationMeier-Stephenson, V., Badmalia, M. D., Mrozowich, T., Lau, K. C. K., Schultz, S. K., Gemmill, D. L., Osiowy, C., van Marle, G., Coffin, C. S., & Patel, T. R. (2021). Identification and characterization of a G-quadruplex structure in the pre-core promoter region of hepatitis B virus covalently closed circular DNA. Journal of Biological Chemistry, 296, Article 100589.en_US
dc.identifier.urihttps://hdl.handle.net/10133/6369
dc.language.isoen_CAen_US
dc.publisherElsevieren_US
dc.publisher.departmentDepartment of Chemistry and Biochemistry
dc.publisher.facultyArts and Science
dc.publisher.institutionUniversity of Lethbridge
dc.publisher.institutionUniversity of Calgary
dc.publisher.institutionPublic Health Agency of Canada
dc.publisher.institutionUniversity of Alberta
dc.publisher.urlhttps://doi.org/10.1016/j.jbc.2021.100589
dc.subjectHepatitis B virusen_US
dc.subjectPre-core promoteren_US
dc.subjectcccDNAen_US
dc.subjectDNA G-quadruplexen_US
dc.subjectHBV surface antigenen_US
dc.subjectELISAen_US
dc.subjectMicroscale thermophoresisen_US
dc.subject.lcshSmall-angle x-ray scattering
dc.titleIdentification and characterization of a G-quadruplex structure in the pre-core promoter region of hepatitis B virus covalently closed circular DNAen_US
dc.typeArticleen_US
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