RAC protein induces enzymatic access to the maturing 3′-end of the 25S rRNA in Schizosaccharomyces pombe

In Schizosaccharomyces pombe, interdependency between steps in the processing of the rRNAs is mediated by a large protein complex (RAC) which interacts with the non-conserved transcribed spacers. The RAC complex exhibits no nuclease activity but dramatically alters the efficiency and specificity of Pac1 nuclease cleavage, leading to the removal of the 3' external transcribed spacers (3'ETS) in the maturation of the 3'ETS region. In this study modification exclusion and S1 nuclease were used to probe the RAC protein binding site and any subsequent structural changes in the maturing region. The results indicate that, as previously observed with the ITS1 and ITS2 regions, the upper helical region in the highly conserved extended terminal hairpin constitutes a protein binding site. In turn, this interaction induces a conformational change which affords access to nuclease at the 3'-end of the maturing 25S rRNA sequence.