Figure 6.

RancRNA_18 cross-links to the 60S E-site region. the sites of cross-linking (x-link) of rancRNA_18 carrying two 4-thio-U modifications (4tU-rancRNA_18) to 80S ribosomes were monitored by primer extension analysis and revealed cross-linking sites to C2675 (red) and C2693 (blue) of helix 84 of the 25S rRNA. experiments in the presence of 4tU-rancRNA_18 without irradiation (lane 4) or in the absence of 4tU-rancRNA_18 with irradiation (lane 5) served as controls. cross-linking experiments and primer extension analyses were performed on two biological replicates. the primer extension stop marked with an asterisk could not always be detected using different primers and thus was excluded from further discussions. Lane 1 (ddTTP) and 2 (ddGTP) indicate dideoxy sequencing lanes. location of cross-linking sites (arrows) are shown in the secondary structure model of yeast 25S rRNA (PTC, peptidyl transferase centre)