Ground squirrel hepatitis virus

Ground squirrel hepatitis virus consists of a DNA genome is primarily double-stranded. Like other hepadnaviruses, it consists of a single-stranded region of variable length that is converted to double-stranded DNA by virion DNA polymerase.[4][6] A protein is covalently bound to the 5' end of the minus strand which may be used as a primer for DNA synthesis.[7]

There are four evident open reading frames within the genome, all on the same strand. Two are for the genes encoding the structural proteins (surface antigens and core antigens), and the other two are unassigned. The larger of the unassigned frames, which is called gene A and covers 80% of the genome, is predicted to encode the viral DNA polymerase. The shorter one, gene B, does not correspond to a known viral protein.

Inside a host cell, the GSHV genome can exist in two forms: a circular duplex and heterogenous partially single-stranded DNA.[6] Circular DNA can be either covalently closed or open.[7]

The GSHV genome is very similar to the related hepadnaviruses. Ground squirrel hepatitis is 3311 base pairs in length, making the size indistinguishable from HBV. GSHV has greater nucleotide and amino acid homology with WHV than HBV, with 82% nucleotide and 78% amino acid homology between GSHV and WHV and 55% nucleotide and 46% amino acid homology between GSHV and HBV.[6][8] The open reading frames of GSHV are similar to HBV and WHV, but the viruses differ in their EcoRI restriction enzyme recognition sites.[4]

Ground squirrel hepatitis virus is a spherical, enveloped virus. The core particle, or nucleocapsid, contains e antigen, viral DNA, and an endogenous DNA polymerase.[9] The virus particles contain two major structural proteins, surface antigens (sAg) and core antigens (cAg). Both proteins were identified based on homology with analogous proteins in other hepadnaviruses. Mature ground squirrel hepatitis sAg (GSHsAg) had 90% amino acid homology with woodchuck hepatitis sAg and is believed to be glycosylated. The ground squirrel hepatitis cAg (GSHcAg) is the most highly conserved protein sequence between GSHV, WHV, and HBV.[6]

It is thought that hepadnaviruses enter host cells through receptor-mediated endocytosis.[2]

The intracellular predomination of the long minus strand of DNA over the short plus strand suggests DNA replication proceeds from an RNA template, like other hepadnaviruses.[6] It is predicted that synthesis of the minus strand utilizes an RNA intermediate that is degraded as it is copied, and the plus strand is synthesized using the complete minus strand DNA. Viral DNA polymerase synthesizes both the plus and minus DNA strands. The production of an RNA intermediate, called the pregenome, during replication involves a reverse transcription step resembling that of retroviruses. Hepadnaviruses differ from retroviruses in that the intermediate form of the genome is RNA instead of DNA. Also, integration into the host genome is not necessary for replication in hepadnaviruses.[2] This process was first shown in Duck Hepatitis B virus, a related virus in the Hepadnaviridae family in which the minus DNA strands also outnumber the plus strands. This similarity suggests the replication mechanism is similar in GSHV.[10]

Following entry, the partial DNA duplex genome is repaired by the viral DNA polymerase and circularized to serve as the template for transcription. The four transcripts are made in the nucleus and are generally transported to the cytoplasm.[2]

It is predicted that viral DNA is packaged before synthesis of the plus strand is complete. This would explain the excess minus strand DNA compared to plus strand DNA.[7]

Ground squirrel hepatitis virus infects liver cells and is considered highly hepatotropic.[1][6] Infection with GSHV can cause persistent infection.[8] The virus causes chronic infection and longer antigenemia (antigen in the blood) when it is injected directly into the liver compared to injection into the portal vein. GSHsAg (viral surface antigen) can be detected in the serum several weeks post infection.[5]

Hepatocellular carcinoma cells

Illness or death in GSHV bearing animals due to liver disease has not been shown. In many infected squirrels with high GSHsAg and virion DNA polymerase levels, there are no signs of cirrhosis or hepatocellular carcinoma.[1] It is possible for ground squirrels persistently infected with GSHV to exhibit signs of mild, nonprogressive hepatitis.[2] No modulation of host processes as a result of GSHV infection is known.

Persistent infection with HBV and WHV often results in hepatocellular carcinoma (HCC). The same association has not been observed with GSHV, but it is possible that persistent infection over a time period longer than required of the other two viruses may contribute to the development of HCC.[8] Persistent infection with GSHV for up to three years only results in slight inflammation, so the development of HCC would presumably take longer in most cases.[6]