Figure 1
From: Chemical inhibitors destabilize HuR binding to the AU-rich element of TNF-α mRNA
Binding affinity of HuR and tristetraprolin to the ARE of TNF-α mRNA. To make fusion proteins, we attached a GST tag to the N-terminal region of HuR (NP_034615) and tristetraprolin (NP_035886). For RNA ligands, we cloned the ARE sequence from TNF-α, COX-2, IL-6, and cFos mRNAs. RRM, RNA-recognition motif; ZnF, Zn2+-finger binding motif. (A) HuR (upper) and tristetraprolin (TTP, lower) bound to the ARE of TNF-α mRNA in RNA EMSA analysis. (B) 32P-labeled TNF-α ARE RNAs were incubated with the indicated concentration of HuR, tristetraprolin or GST protein. After incubation at 25℃ for 20 min, the reaction mixture was resolved by gel electrophoresis in a 6% PAGE gel. (C) The affinity of HuR, tristetraprolin, and GST for TNF-α was determined by filter binding assay.
