Figure 5
From: Cancer-associated noncoding mutations affect RNA G-quadruplex-mediated regulation of gene expression
Two adjacent mutations in the 5′ UTR of CXCL14 destabilise the RNA G4 and affect its regulatory function. (A) Schematic representation of three different mRNAs of firefly luciferase reporter constructs. Wild-type construct contains full-length 5′ UTR of CXCL14. In the G4-mutated construct, two guanines are replaced by two adenines. To make the G4-deleted construct, 23 nucleotides that predicted to form the RNA 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 CXCL14 wild-type and mutated RNA G4 oligos in KCl. (C) CD melting curves of CXCL14 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).
