Hepatitis B virus HBx protein localized to the nucleus restores HBx-deficient virus replication in HepG2 cells and in vivo in hydrodynamically-injected mice

Abstract Identifying the requirements for the regulatory HBx protein in hepatitis B virus (HBV) replication is an important goal. A plasmid-based HBV replication assay was used to evaluate whether HBx subcellular localization influences its ability to promote virus replication, as measured by real t...

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Bibliographic Details
Published in:Virology (New York, N.Y.) N.Y.), 2009-07, Vol.390 (1), p.122-129
Main Authors: Keasler, Victor V, Hodgson, Amanda J, Madden, Charles R, Slagle, Betty L
Format: Article
Language:English
Subjects:
60 APPLIED LIFE SCIENCES
ANIMALS
Base Sequence
Cell Line
Cell Nucleus - virology
DIGESTIVE SYSTEM DISEASES
DISEASES
DNA Primers - genetics
DNA, Viral - genetics
DNA, Viral - metabolism
Female
GENE AMPLIFICATION
Gene Expression
Genes, Viral
Genetic Vectors
HBV replication
HBx
HEPATITIS
Hepatitis B virus
Hepatitis B virus - genetics
Hepatitis B virus - physiology
Humans
Infectious Disease
INJECTION
INTAKE
MAMMALS
MICE
Mice, Inbred ICR
MICROORGANISMS
Nuclear Export Signals - genetics
Nuclear localization
Nuclear Localization Signals - genetics
ORGANIC COMPOUNDS
PARASITES
POLYMERASE CHAIN REACTION
PROTEINS
RADIATION, THERMAL, AND OTHER ENVIRONMENTAL POLLUTANT EFFECTS ON LIVING ORGANISMS AND BIOLOGICAL MATERIALS
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
RODENTS
Trans-Activators - genetics
Trans-Activators - physiology
VERTEBRATES
Virus Replication - physiology
VIRUSES
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Summary:Abstract Identifying the requirements for the regulatory HBx protein in hepatitis B virus (HBV) replication is an important goal. A plasmid-based HBV replication assay was used to evaluate whether HBx subcellular localization influences its ability to promote virus replication, as measured by real time PCR quantitation of viral capsid-associated DNA. HBx targeted to the nucleus by a nuclear localization signal (NLS-HBx) was able to restore HBx-deficient HBV replication, while HBx containing a nuclear export signal (NES-HBx) was not. Both NLS-HBx and NES-HBx were expressed at similar levels (by immunoprecipitation and Western blotting), and proper localization of the signal sequence-tagged proteins was confirmed by deconvolution microscopy using HBx, NLS-HBx, and NES-HBx proteins fused to GFP. Importantly, these findings were confirmed in vivo by hydrodynamic injection into mice. Our results demonstrate that in these HBV replication assays, at least one function of HBx requires its localization to the nucleus.
ISSN:0042-6822
1096-0341
DOI:10.1016/j.virol.2009.05.001