ABSTRACT
Arenaviruses and filoviruses are capable of causing hemorrhagic fever syndrome in humans. Limited therapeutic and/or prophylactic options are available for humans suffering from viral hemorrhagic fever. In this report, we demonstrate that pre-treatment of host cells with the kinase inhibitors genistein and tyrphostin AG1478 leads to inhibition of infection or transduction in cells infected with Ebola virus, Marburg virus, and Lassa virus. In all, the results demonstrate that a kinase inhibitor cocktail consisting of genistein and tyrphostin AG1478 is a broad-spectrum antiviral that may be used as a therapeutic or prophylactic against arenavirus and filovirus hemorrhagic fever.
Subject(s)
Antiviral Agents/pharmacology , Ebolavirus/drug effects , Genistein/pharmacology , Hemorrhagic Fever, Ebola/virology , Lassa Fever/virology , Lassa virus/drug effects , Protein Kinase Inhibitors/pharmacology , Quinazolines/pharmacology , Tyrphostins/pharmacology , Cell Line , Ebolavirus/genetics , Ebolavirus/physiology , Hemorrhagic Fever, Ebola/drug therapy , Humans , Lassa Fever/drug therapy , Lassa virus/genetics , Lassa virus/physiologySubject(s)
Edetic Acid/analogs & derivatives , Edetic Acid/analysis , Electrophoresis, Polyacrylamide Gel/methods , Iron Chelating Agents/analysis , Iron Radioisotopes/analysis , Organometallic Compounds/analysis , Sulfhydryl Compounds/analysis , Viral Proteins/analysis , Amino Acid Substitution , Cysteine/chemistry , Electrophoretic Mobility Shift Assay , Isotope Labeling , Oxidation-Reduction , Viral Proteins/chemistryABSTRACT
VP39 is a vaccinia virus-encoded RNA modifying protein with roles in the modification of both mRNA ends. At the 3' end it acts as a processivity factor for the vaccinia poly(A) polymerase (VP55), promoting poly(A) tail elongation. Despite VP39's three-dimensional structure having been elucidated along with details of its mode of mRNA 5' end binding, the VP39-VP55 heterodimer's molecular mechanism of processivity is largely unknown. Here, the area immediately above almost the entire surface of the VP39 subunit was probed using chemical reporters, and the path of a previously unidentified RNA binding site was revealed. The path was indicated to fall within a cleft formed by the intersubunit interface and was consistent with both a previously reported model of the heterodimer-nucleic acid ternary complex and the known function of the heterodimer in processive poly(A) tail elongation.
