FIG. 5.

Transcripts whose 3′ end is generated by a self-cleaving hammerhead ribozyme accumulate in Rrp6p-dependent transcription site foci. (A) GFP RNA FISH analysis of wild-type (top row) and Δrrp6 (middle row) strains transformed with a 2μm high-copy-number plasmid construct expressing GFP RNA terminated by a self-cleaving hammerhead ribozyme (polII-GFP-RZ) and wild-type cells transformed with a 2μm plasmid expressing GFP RNA terminated by a conventional 3′-end processing signal (polII-GFP) (bottom row). Δrrp6 cells were cotransformed with either a plasmid expressing the Rrp6p protein (top and bottom rows) or empty vector (middle row). Log-phase cultures growing at 30°C were fixed and subjected to RNA FISH utilizing probes specific for the GFP ORF. DNA was stained with DAPI. dT, oligo(dT). (B) Northern analysis of GFP RNA from the cultures described above. WT, wild type.