Figure 8: In vitro cytidine deamination of SDHB RNA and ssDNA by APOBEC3A
From: APOBEC3A cytidine deaminase induces RNA editing in monocytes and macrophages
(a) c.136C>U editing of an ∼1.1 kb exogenous SDHB ORF RNA by whole-cell lysates of control or APOBEC3A 293T transfectants. Duration of the deamination reactions and amount of lysate protein in them are noted. For some reactions, lysates were pre-heated at 85 °C for 15 min. (b) c.136C>U editing of the RNA by 10 μM purified C-His6-tagged APOBEC3A protein. The reactions had 180 amole SDHB RNA and 100 nM ZnCl2. (c) Sanger sequence chromatogram traces of PCR amplified products of SDHB deamination reactions that had either 180 amole of ∼1.1 kb SDHB RNA or 100 amole of SDHB ssDNA of 120 b as substrate. APOBEC3A protein was present in the+reactions at 5 and 20 μM in the reaction with RNA and DNA substrate, respectively. Reactions for b and c conducted for two hours at 37 °C. Mean and range (n=3) are shown in a and b, respectively.
