Figure 3
From: Cancer-associated noncoding mutations affect RNA G-quadruplex-mediated regulation of gene expression
A point mutation in the 5′ UTR of BCL2 destabilises the RNA G4 and affects its regulatory function. (A) Schematic representation of three different mRNAs of firefly luciferase reporter constructs. Wild-type construct contains full-length 5′ UTR of BCL2. In the G4-mutated construct, one of the guanines is replaced by adenine. To make the G4-deleted construct, 22 nucleotides that form the G4 were deleted. Schematic drawing of wild-type and mutated G4s are proposed based on RNAfold and QGRS-mapper predictions and in vitro results. (B) CD spectra of BCL2 wild-type and mutated RNA G4 oligos in KCl. (C) CD melting curves of BCL2 wild-type and mutated RNA G4 oligos at 260 nm. (D) Relative in vitro translation efficiency of the wild-type, G4-mutated and G4-deleted constructs as judged by measuring firefly luciferase enzymatic activity. Data were analysed using one-way-ANOVA (n ≥ 3, error bars represent the SEM, asterisks (*) indicate significance calculated by the Tukey multiple comparisons test, ns indicates non-significant).
