AID	Name	Type	Comment
588638	qHTS for Inhibitors of Polymerase Kappa: Summary	summary

The processing and repair of DNA interstrand crosslinks (ICL) in eukaryotic cells are extremely complex, potentially involving multiple DNA repair and damage tolerance pathways, including homologous recombination, nucleotide excision repair, translesion synthesis (TLS), and cell cycle checkpoints. In some repair models, the dually-incised strand possesses sufficient mobility that a DNA polymerase can strand displace the nucleotide patch that is 5' to the lesion, then replicate past the ICL site to complete the repair gap-filling synthesis. Polymerase kappa has been implicated in this project and is hence of interest.

To that end, in a collaboration between the Oregon Health & Science University and the NIH Chemical Genomics Center a 1536-well high-throughput screen was developed. This fluorescent screen was assayed against the NIH Molecular Libraries Small Molecule Repository (MLSMR) to discover inhibitors of Pol Kappa.


NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]

MLPCN Grant: MH094179
Assay Submitter (PI): Stephen Lloyd, Oregon Health & Science University

Three microliters of reagents (buffer in column 3 and 4 as negative control and 10 nM Pol kappa in columns 1, 2, and 5-48) were dispensed into a 1536-well black solid-bottom plate. Compounds (23 nL) were transferred via Kalypsys pin tool equipped with 1536-pin array. The plates were then incubated for 15 min at room temperature, and 1 uL substrate (50 nM final concentration) were then added to start the reaction and kinetically read twice at 0 min and 10 min on the Viewlux reader

Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.

Grant Number: MH094179