AID	Name	Type	Comment
624266	qHTS for Small Molecule Inhibitors of the ERG Ets/DNA interaction: Summary.	summary

The Ets family of proteins is a family of ~30 conserved transcription factors defined by the presence of an ~85 amino acid domain referred to as the Ets domain, which mediates sequence-specific DNA binding to a core DNA element GGAA/T.The Ets family member ERG has been linked to several neoplasms. ERG has been shown to be frequently over-expressed in prostate cancer. Perhaps more strikingly, ERG and the Ets protein ETV1 have recently been shown to be the targets of chromosomal translocations with TMPRSS2 which are observed in 80% of prostate cancer patient samples. More extensive studies of the effects of over-expression of ERG in prostate cells have recently been carried out and have shown that over-expression of ERG results in increased invasion.

The importance of DNA-binding to the function of these proteins strongly suggests that inhibiting the DNA binding activity of ERG may be a mechanism to treat prostate cancer as well as other cancers where the activity of ERG plays a key role. We will carry out a high throughout screen to identify inhibitors of the interaction between ERG and DNA using a fluorescence polarization (FP) assay with a His-tagged ERG Ets domain and fluorescently-labeled DNA probes to develop small molecule inhibitors of this interaction as probes to test this hypothesis and lay the groundwork for the development of targeted therapies against ERG. This is the confirmation against the cherry picks in theprimary screening assay.

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

Grant: NS066464
PI Name: John Bushweller, University of Virgina

1.5 ul of protein solution (1 uM ERG domain in 300mM KCl, 50mM Tris-HCl, 1mM EDTA, 1mM DTT, pH 7.5); 20 nl Control plate and compound library plate; 1.5 ul mixture of 5 nM Fluorescein-DNA and 100nM Texas-615-DNA solution in same buffer as above solution; measure fluorescence polarization sequentially for fluorescein (Excitation = 480 nm, Emission = 535s, 535p nm) and Texas red (Excitation = 555 nm, Emission = 632s, 632p nm).

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: NS066464