The effects of L7Ae, catalytic aspartate, and Nop10 on substrate RNA docking. Fluorescence intensity at 363 nm was monitored while a substrate RNA labeled with both 2-AP and 5-FU and bound with guide RNA (gRNA), Cbf5, Nop10, and Gar1 (CNG RNP) was titrated with saturating amounts of L7Ae (L). Comparison of the fluorescence intensity profiles of L7Ae titration to the wild-type CNG RNP (A), L7Ae titration to the CGN RNP containing catalytic aspartate to alanine mutation (CmNG) (B), L7Ae titration to CNG RNP containing Nop10 Arg34 to alanine mutation (CNmG) (C). Labels “1,” “2,” and “3” indicate the order of titration steps of the specified subunits. Arrows indicate titration points.
