High-throughput screening assay for helicase enzymes.
We have developed a novel 96-well microtiter plate high-throughput screening filtration assay for the detection of helicase activity. In this paper we present data for detection of helicase activity of the UL5/8/52 helicase-primase complex from herpes simplex virus 1 (HSV1). The assay involves the detection of radioactively labeled oligonucleotide annealed to a single-stranded circular DNA following capture of the annealed complex on silica beads. We have screened over 200,000 samples containing small organic molecules and natural products and identified T157602, a two-amino thiazole, as a specific inhibitor of HSV replication.
Sivaraja
M
M
Tularik Inc., South San Francisco, California 94080, USA. mohan@Tularik.com
Giordano
H
H
Peterson
M G
MG
D001483:Base Sequence; D002021:Buffers; D004265:DNA Helicases / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism; D019915:DNA Primase; D017931:DNA Primers; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018259:Herpesvirus 1, Human / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D012371:Robotics; D014764:Viral Proteins
D002021:Buffers; D017931:DNA Primers; D004791:Enzyme Inhibitors; D014764:Viral Proteins; D019915:DNA Primase; C062021:helicase-primase, Human herpesvirus 1; D004265:DNA Helicases