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The hepatitis B virus-associated reverse transcriptase is encoded by the viral pol gene
M Bavand1, M Feitelson, O Laub
1Department of Genetics, Weizmann Institute of Science, Rehovot, Israel.
Journal of Virology
|February 1, 1989
Summary
Hepatitis B virus (HBV) reverse transcriptase activity was detected in HBV particles. This activity is linked to the polymerase (Pol)-open reading frame (ORF), suggesting HBV encodes its own reverse transcriptase.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Hepatitis B virus (HBV) is a significant human pathogen.
- The genetic origin of HBV reverse transcriptase has remained unclear.
- Identifying the source of viral enzymes is crucial for understanding viral replication.
Purpose of the Study:
- To investigate the molecular identity of the reverse transcriptase activity associated with HBV.
- To determine if the polymerase (Pol)-open reading frame (ORF) encodes the viral reverse transcriptase.
Main Methods:
- Activity gel electrophoresis to detect enzymatic activity in HBV particles.
- Western immunoblotting using specific antisera to identify viral proteins.
- Immunoprecipitation assays to confirm protein-enzyme interactions.
Main Results:
- Major (approx. 70 kDa) and minor (approx. 90 kDa) reverse transcriptase activities were found in infectious HBV particles.
- A 70-kDa peptide recognized by antisera against the carboxy terminus of the Pol protein was detected in HBV particles and cell extracts.
- Eluted reverse transcriptase protein from activity gels reacted with Pol-specific antisera, and activity was immunoprecipitated by these antisera.
Conclusions:
- The findings provide strong evidence that the HBV polymerase (Pol)-ORF encodes the viral reverse transcriptase.
- This discovery advances the understanding of HBV replication mechanisms.
- It suggests that HBV possesses its own enzymatic machinery for replication.