3,7-Dihydroxytropolones Inhibit Initiation of Hepatitis B Virus Minus-Strand DNA Synthesis

Author:

Bak, Ellen

Miller,Jennifer

Noronha, Andrea

Tavis, John

Gallicchio, Emilio

Murelli, Ryan P

Le Grice,Stuart
Author Address

Basic Research Laboratory National Cancer Institute, Frederick, MD 21702, USA., Department of Molecular Microbiology and Immunology, St. Louis University, St. Louis, MO 63104, USA., Department of Chemistry, Brooklyn College, The City University of New York, Brooklyn, NY 11210, USA., PhD Program in Chemistry, The Graduate Center of The City University of New York, New York, NY 10016, USA., PhD Program in Biochemistry, The Graduate Center of The City University of New York, New York, NY 10016, USA.,

Abstract:

Initiation of protein-primed (-) strand DNA synthesis in hepatitis B virus (HBV) requires interaction of the viral reverse transcriptase with epsilon (e), a cis-acting regulatory signal located at the 5 39; terminus of pre-genomic RNA (pgRNA), and several host-encoded chaperone proteins. Binding of the viral polymerase (P protein) to e is necessary for pgRNA encapsidation and synthesis of a short primer covalently attached to its terminal domain. Although we identified small molecules that recognize HBV e RNA, these failed to inhibit protein-primed DNA synthesis. However, since initiation of HBV (-) strand DNA synthesis occurs within a complex of viral and host components (e.g., Hsp90, DDX3 and APOBEC3G), we considered an alternative therapeutic strategy of allosteric inhibition by disrupting the initiation complex or modifying its topology. To this end, we show here that 3,7-dihydroxytropolones (3,7-dHTs) can inhibit HBV protein-primed DNA synthesis. Since DNA polymerase activity of a ribonuclease (RNase H)-deficient HBV reverse transcriptase that otherwise retains DNA polymerase function is also abrogated, this eliminates direct involvement of RNase (ribonuclease) H activity of HBV reverse transcriptase and supports the notion that the HBV initiation complex might be therapeutically targeted. Modeling studies also provide a rationale for preferential activity of 3,7-dHTs over structurally related a-hydroxytropolones (a-HTs).

See More

Author Address