Biochemical and structural bioinformatics studies of fungal CutA nucleotidyltransferases explain their unusual specificity toward CTP and increased tendency for cytidine incorporation at the 3′-terminal positions of synthesized tails

Substitution of several amino acids within Cid1 NRM increases its specificity toward CTP. (A) Length distribution for tails added by wild-type (WT) and D330E/E333N/H336R triple mutant (PCP) Cid1 variants to ss22-A₄ (left) and ss22-U₄ (right) oligoribonucleotide substrates. Numbers of analyzed sequences, obtained after 3′-RACE-seq high-throughput analysis for each protein variant, are indicated in parentheses. (B,C) Analysis of nucleotide composition for tails added by Cid1 WT and PCP variants to ss22-A₄ (panel B) and ss22-U₄ (panel C) RNA substrates. Results are presented as in Figure 6.