Figure 8.

Rrp6–TAP co-purifies early precursor rRNAs. A, Δnop53/tetOff::GFP-NOP53/RRP6-TAP strain was used for RNA coimmunoprecipitation with Rrp6–TAP in the presence (−Dox (doxycycline )), or absence (+Dox) of Nop53. Total RNA extracted from Δrrp6 strain was used as a control of precursor rRNAs accumulating in the absence of Rrp6. Left panels show two biological replicates of RNAs extracted from aliquots of total cell extracts in the indicated conditions used in the coimmunoprecipitations as shown on the right panels. Precursor rRNAs are indicated on the right. 5S rRNA and scR1 were used as controls for nonspecific binding to the resin. Total RNA extracted from replicate 1 in the absence of Nop53 (+Dox) was lost during loading on the gel, but it was maintained in the figure because it clearly shows the accumulation of 7S pre-rRNA under this condition. B, schematic representation of the yeast 35S pre-rRNA indicating the hybridizing positions of the different probes.