Figure 4.
Applying borohydride-dependent misincorporation to study ac4C in rRNA biogenesis. (a) RNA modifications in human 18S helix 45. (b) RT of 21S-C pre-rRNA depends on relative timing of m26A and ac4C modifications. “C” and “E” represent sites at which the pre-rRNA is cleaved. Extension of the 21S-C primer, which forms cDNA from pre-rRNAs in which these cleavage sites are intact, may be impeded by m26A and reduced ac4C. (c) ac4C-dependent mismatch can be detected using an RT primer specific for the 21S-C pre-rRNA and its precursors.