Identification and Validation of Novel Small Molecule Disruptors of HuR-mRNA Interaction

• DOI: 10.1021/cb500851u
• Publication Date: 2015-03-09T19:07:38Z
• Authors: Xiaoqing Wu; Lan Lan; David Michael Wilson; Rebecca T. Marquez; Wei-chung Tsao; Philip Gao; Anuradha Roy; Benjamin Andrew T...; Peter McDonald; Jon A Tunge; Steven A Rogers; Dan A. Dixon; Jeffrey Aubé; Liang Xu
• Keywords: Immunoprecipitation

HuR, an RNA binding protein, binds to adenine- and uridine-rich elements (ARE) in the 3'-untranslated region (UTR) of target mRNAs, regulating their stability translation. HuR is highly abundant many types cancer, it promotes tumorigenesis by interacting with cancer-associated which encode proteins that are implicated different tumor processes including cell proliferation, survival, angiogenesis, invasion, metastasis. Drugs disrupt stabilizing effect upon mRNA targets could have dramatic effects on inhibiting cancer growth persistence. In order identify small molecules directly HuR-ARE interaction, we established a fluorescence polarization (FP) assay optimized for high throughput screening (HTS) using protein ARE oligo from Musashi RNA-binding 1 (Msi1) mRNA, target. Following performance HTS ∼6000 compounds, discovered cluster potential disruptors, were then validated AlphaLISA (Amplified Luminescent Proximity Homogeneous Assay), surface plasmon resonance (SPR), ribonucleoprotein immunoprecipitation (RNP IP) assay, luciferase reporter functional studies. These compounds disrupted interactions at nanomolar level blocked function competitive HuR. results support future studies toward chemical probes study possibly novel therapy HuR-overexpressing cancers.