Dataset Information


Cell-Based Ligand Discovery for the ENL YEATS Domain.

ABSTRACT: ENL is a transcriptional coactivator that recruits elongation machinery to active cis-regulatory elements upon binding of its YEATS domain-a chromatin reader module-to acylated lysine side chains. Discovery chemistry for the ENL YEATS domain is highly motivated by its significance in acute leukemia pathophysiology, but cell-based assays able to support large-scale screening or hit validation efforts do not presently exist. Here, we report on the discovery of a target engagement assay that allows for high-throughput ligand discovery in living cells. This assay is based on the cellular thermal shift assay (CETSA) but does not require exposing cells to elevated temperatures, as small-molecule ligands are able to stabilize the ENL YEATS domain at 37 °C. By eliminating temperature shifts, we developed a simplified target engagement assay that requires just two steps: drug treatment and luminescence detection. To demonstrate its value for higher throughput applications, we miniaturized the assay to a 1536-well format and screened 37 120 small molecules, ultimately identifying an acyl-lysine-competitive ENL/AF9 YEATS domain inhibitor.

PROVIDER: S-EPMC7384521 | BioStudies |

REPOSITORIES: biostudies

Similar Datasets

| S-EPMC8161486 | BioStudies
| S-EPMC5497220 | BioStudies
| S-EPMC6348381 | BioStudies
| S-EPMC6503841 | BioStudies
| S-EPMC5372383 | BioStudies
| S-EPMC5828395 | BioStudies
| S-EPMC4871749 | BioStudies
2017-02-23 | GSE82117 | GEO
2017-02-23 | GSE82116 | GEO
| S-EPMC5014688 | BioStudies