Fig. 4: PERK regulates the structural changes of human p53 mRNA under ER stress.

a DMS-footprint of human p53 mRNA under normal, ER stress and ER stress with PERKΔC over-expression. The presence of the dominant-negative PERKΔC reverses the methylation pattern, indicating that the changes in the p53 mRNA structure following ER stress are PERK-dependent. Modifications reversed by PERKΔC are marked with red boxes. b ΔSHAPE analysis demonstrating significant structural alterations in p53 mRNA following the ER stress (Thap), with RNA regions constrained during the stress are highlighted in green and the regions opened or exposed in violet. c ΔSHAPE analysis demonstrating that over-expression of dominant-negative mutant PERKΔC reverses the structural alterations generated during ER stress. d ΔSHAPE analysis of mouse p53 mRNA did not show structural alterations following the ER stress. e Top panel: cartoon depicting human and mouse p53-WT constructs with a short ORF inserted upstream of the 1st AUG. Bottom panel: Western blot showing the expression of p53 and p47/p44 isoforms with the indicated constructs.