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Detection of Low Copy Number Integrated Viral DNA Formed by In Vitro Hepatitis B Infection
Published on: November 7, 2018
Maturation-associated destabilization of hepatitis B virus nucleocapsid
Xiuji Cui1, Laurie Ludgate, Xiaojun Ning
1Department of Microbiology and Immunology, Pennsylvania State University College of Medicine, Hershey, Pennsylvania, USA.
Journal of Virology
|August 23, 2013
Summary
Hepatitis B virus nucleocapsids (NCs) undergo maturation, becoming destabilized and structurally changed. This maturation selectively targets NCs for either virion secretion or DNA uncoating, crucial for viral replication.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Hepatitis B virus (HBV) nucleocapsids (NCs) containing relaxed circular (RC) DNA are essential for viral replication and persistence.
- Mature NCs are involved in either extracellular virion secretion or nuclear uncoating to form covalently closed circular (CCC) DNA.
- Immature NCs are not competent for these processes, but the selection mechanism for mature NCs remains unclear.
Purpose of the Study:
- To biochemically analyze different populations of HBV nucleocapsids (NCs).
- To understand the structural changes associated with NC maturation.
- To elucidate the role of these changes in the selective processing of mature NCs.
Main Methods:
- Separation of nucleocapsid (NC) populations using sucrose gradient centrifugation.
- Biochemical analysis including protease and nuclease sensitivity assays.
- Native agarose gel electrophoresis to assess NC mobility.
- In vitro generation of mature NCs for comparative analysis.
Main Results:
- NC maturation is associated with destabilization, evidenced by increased protease and nuclease sensitivity.
- Mature NCs exhibit altered sedimentation properties and retarded mobility during native agarose gel electrophoresis.
- Three distinct populations of intracellular mature NCs were identified based on these characteristics.
- In vitro generated mature NCs displayed similar destabilized properties.
Conclusions:
- Significant structural changes occur during HBV nucleocapsid (NC) maturation, leading to destabilization.
- These structural alterations are likely key to the selective uncoating of mature NCs for CCC DNA formation.
- Destabilization also facilitates the preferential envelopment of mature NCs for virion secretion.
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