Transcription of ribosomal DNA by RNA polymerase I (Pol I) requires the
initiation factor Rrn3. Here we report the cryo-EM structure of the Pol I-Rrn3
complex at 4.8 Å resolution. The structure reveals how Rrn3 binding converts an
inactive Pol I dimer into an initiation-competent monomeric complex and provides
insights into the mechanisms of Pol I-specific initiation and regulation.