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BioAssay: AID 588591

qHTS for Inhibitors of Polymerase Eta

The Y-family of DNA polymerases, such as Pol eta, are specifically involved in DNA repair. Pol eta copies undamaged DNA with a lower fidelity than other DNA-directed polymerases. However, it accurately replicates UV-damaged DNA; when thymine dimers are present, this polymerase inserts the complementary nucleotides in the newly synthesized DNA, thereby bypassing the lesion and suppressing the more ..
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 Tested Compounds
 Tested Compounds
All(388711)
 
 
Active(4729)
 
 
Inactive(370338)
 
 
Inconclusive(13864)
 
 
 Tested Substances
 Tested Substances
All(392343)
 
 
Active(4746)
 
 
Inactive(373704)
 
 
Inconclusive(13893)
 
 
AID: 588591
Data Source: NCGC (PolE100)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-10-20

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 4729
Related Experiments
AIDNameTypeComment
588636qHTS for Inhibitors of Polymerase Eta: SummarySummarydepositor-specified cross reference
720502qHTS for Inhibitors of Polymerase Eta: Confirmatory Assay for Cherry-picked CompoundsConfirmatorydepositor-specified cross reference
Description:
The Y-family of DNA polymerases, such as Pol eta, are specifically involved in DNA repair. Pol eta copies undamaged DNA with a lower fidelity than other DNA-directed polymerases. However, it accurately replicates UV-damaged DNA; when thymine dimers are present, this polymerase inserts the complementary nucleotides in the newly synthesized DNA, thereby bypassing the lesion and suppressing the mutagenic effect of UV-induced DNA damage. Pol eta has the ability to bypass cisplatinated DNA adducts in vitro, and it has been suggested that pol eta-dependent bypass of the cisplatin lesion in vivo leads to increased tumor resistance. Thus, while pol eta's (and most likely iota's) normal function is to protect humans against the deleterious consequences of DNA damage, under certain conditions they can have deleterious effects on human health. As a consequence, we propose to utilize a high throughput replication assay to identify small molecule inhibitors of pol eta.

In a collaboration between the National Institute of Child Health & Human Development (NICHD) and NIH Chemical Genomics Center, a high-throughput, fluorescent screen was developed to screen the NIH Molecular Libraries Small Molecule Repository (MLSMR). This screen is used to identify inhibitors of pol eta.

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

MLPCN Grant: MH090825
Assay Submitter (PI): Roger Woodgate, NICHD)
Protocol
Three microliters of reagents (buffer in column 3 and 4 as negative control and 10 nM Pol eta in columns 1, 2, and 5-48) were dispensed into a 1,536-well black solid-bottomed 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 muL substrate (50 nM final concentration) was added to start the reaction and kinetically read twice at 0 min and 10 min on the Viewlux reader
Comment
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.
Categorized Comment - additional comments and annotations
From ChEMBL:
Assay Type: Functional
Assay Cell Type: NULL
Result Definitions
Show more
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
6Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
7Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
8Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
9Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
10Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
11Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
13Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14Activity at 0.00366 uM (0.00366μM**)% Activity at given concentration.Float%
15Activity at 0.018 uM (0.0183μM**)% Activity at given concentration.Float%
16Activity at 0.091 uM (0.0914μM**)% Activity at given concentration.Float%
17Activity at 0.457 uM (0.457μM**)% Activity at given concentration.Float%
18Activity at 2.290 uM (2.29μM**)% Activity at given concentration.Float%
19Activity at 11.40 uM (11.4μM**)% Activity at given concentration.Float%
20Activity at 57.10 uM (57.1μM**)% Activity at given concentration.Float%
21Activity at 114.0 uM (114μM**)% Activity at given concentration.Float%
22Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH090825

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
Classification
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