High-resolution mapping of transcribing Pol III by CRAC in the WT and C128 mutants

Here we report high-resolution analyses of transcribing RNA polymerase III (Pol III) in a wild-type (WT) strain and three C128 mutants. C128 is the second largest subunit of Pol III, which is the key subunit to recognize termination signal. Mutating key residues in C128 interferes with transcription termination in vitro and affect yeast viability as well, raising the question that whether these mutations provoke Pol III termination defects in vivo. Here we show that C128 mutants display termination defects at most PolIII-dependent genes, indicating that the most disruptive C128 mutations indeed impair Pol III transcription termination in vivo Analysis of the effect of the mutations of C128 on Pol III-dependent transcription genome-wide using CRAC