Budihas SR, Beutler JA, Parniak MA, Crouch RJ, Wamiru A, Gardella R, Wilson J, Gaidamakov S, Shulley N, Chan K, Le Grice SF, McMahon JB; HIV DRP Symposium Antiviral Drug Resistance (4th: 2003: Chantilly, Va.).
Program Abstr HIV DRP Symp Antivir Drug Resist. 2003 Dec 7-10; 4: Abstract no. 49.
HIV Drug Resistance Program, Center for Cancer Research, NCI-Frederick, Frederick, MD
Human immunodeficiency virus-type 1 (HIV-1) reverse transcriptase (RT) is a dual-function enzyme that catalyzes DNA polymerization and ribonuclease H (RNase H) activities. RNase H specifically hydrolyzes the RNA strand of an RNA/DNA hybrid. While both activities are critical for infectivity, only the polymerase activity has been a successful target for commercial drugs. Our laboratories are involved in a screening campaign to look for compounds that inhibit HIV-1 RNase H activity. We are using a rapid fluorescence-based assay to screen small-molecule libraries. Due to the nature of the primary screening assay, we could not eliminate from our hits small molecules that appeared to inhibit due to quenching of the fluorescein signal. This was of particular concern for the screening of natural product extracts. Our solution was the development of a rapid fluorescence-based capillary electrophoresis assay that allowed for simultaneous quantitative and qualitative analyses in a moderate throughput format. Multiple chemical libraries totaling nearly 190,000 samples were screened during this campaign. Hits from the initial screens were confirmed and processed through our capillary electrophoresis, dose-response enzyme panel and cytopathicity secondary screens. Several compounds have been identified as leads for medicinal chemistry and further drug development. In addition, we have identified several compounds that may prove to be useful biochemical probes for the study of RT enzymology as well as the human and E. coli RNases H.
Publication Types:
Keywords:
- DNA Primers
- Electrophoresis, Capillary
- HIV-1
- Humans
- RNA
- RNA Probes
- RNA-Directed DNA Polymerase
- Ribonuclease H
- antagonists & inhibitors
- chemistry
- enzymology
- genetics
- metabolism
Other ID:
UI: 102268474
From Meeting Abstracts