An in vitro fluorescence screen to identify antivirals that disrupt hepatitis B virus capsid assembly.
Virus assembly has not been routinely targeted in the development of antiviral drugs, in part because of the lack of tractable methods for screening in vitro. We have developed an in vitro assay of hepatitis B virus (HBV) capsid assembly, based on fluorescence quenching of dye-labeled capsid protein, for testing potential inhibitors. This assay is adaptable to high-throughput screening and can identify small-molecule inhibitors of virus assembly that prevent, inappropriately accelerate and/or misdirect capsid formation to yield aberrant particles. An in vitro primary screen has the advantage of identifying promising lead compounds affecting assembly without the requirement that they be taken up by cells in culture and be nontoxic. Our approach may facilitate the identification of antivirals targeting viruses other than HBV, such as avian influenza and HIV.
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, 975 NE 10th St. BRC 456, Oklahoma City, Oklahoma 73104, USA.
Stephen J
SJ
Stray
Jennifer M
JM
Johnson
Benjamin G
BG
Kopek
Adam
A
Zlotnick
D000998:Antiviral Agents / Q000494:pharmacology*; D001681:Biological Assay; D001896:Boron Compounds; D002213:Capsid / Q000737:chemistry / Q000187:drug effects*; D036022:Capsid Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D019281:Dimerization; D005453:Fluorescence; D005456:Fluorescent Dyes; D006515:Hepatitis B virus / Q000737:chemistry / Q000187:drug effects*; D066298:In Vitro Techniques; D007700:Kinetics; D008027:Light; D008958:Models, Molecular; D012542:Scattering, Radiation; D019065:Virus Assembly / Q000187:drug effects*
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D000998:Antiviral Agents; D001896:Boron Compounds; D036022:Capsid Proteins; D005456:Fluorescent Dyes