Expression, purification, and characterization of an active RNase H domain of the hepatitis B viral polymerase.

The Journal of Biological Chemistry
X Wei, D L Peterson

Abstract

The replication of the hepatitis B viral DNA genome proceeds through a pregenomic RNA intermediate. This pregenomic RNA subsequently serves as the template for the formation of the viral DNA by the reverse transcriptase activity of the viral P gene product. The P gene product is believed to be a multifunctional enzyme with DNA-dependent DNA polymerase, RNA-dependent DNA polymerase, and RNase H activities. Detailed biochemical studies of this protein have not been performed because of the inability to obtain sufficient amounts of the enzyme from the virus and by the inability to produce the enzyme in heterologous expression systems. The RNase H activity is essential for viral replication and is believed to be responsible for the degradation of the RNA pregenomic intermediate as well as for generating the short RNA primer that is required for DNA second strand synthesis. We have assembled an expression vector which directs the synthesis of a protein that corresponds to the putative RNase H domain of the P gene product and having a carboxyl-terminal polyhistidine tag to facilitate purification. The protein has been expressed in Escherichia coli and purified to yield 1-2 mg of protein/liter of culture. This protein has RNase H acti...Continue Reading

References

Jan 1, 1990·Pharmacology & Therapeutics·U Wintersberger
Mar 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·N Tanese, S P Goff
Jan 1, 1987·Annual Review of Biochemistry·D Ganem, H E Varmus
Sep 1, 1981·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·W SzmunessA Kellner
Sep 1, 1995·Journal of Virology·S M Oberhaus, J E Newbold

❮ Previous
Next ❯

Citations

Sep 25, 1999·Journal of Cellular Physiology·M A Feitelson
Jul 30, 2008·Journal of the American Chemical Society·Marco De VivoMichael L Klein
Dec 4, 2010·World Journal of Gastroenterology : WJG·Yang YuSeong-Tshool Hong
May 31, 2014·World Journal of Gastroenterology : WJG·Ceen-Ming TangJun Yu
Apr 12, 2015·Antiviral Research·John E Tavis, Elena Lomonosova
May 18, 2007·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Robert A HoltDuane E Smailus

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.