Pre-ribosomal RNA is processed in permeabilised cells at the site of transcription

The available data concerning the subnucleolar localisation of the individual steps of precursor-ribosomal RNA (pre-rRNA) processing are ambiguous. According to in situ hybridisation studies, the late steps of pre-rRNA processing have been located into the granular component of the nucleolus, but factors engaged in these events were found being enriched in the dense fibrillar component. In this study, by utilisation of permeabilised human cells, we demonstrate that the newly synthesised, bromouridine-labelled pre-rRNAs reside at, or near, the sites of transcription. We provide evidence that processing of pre-rRNA occurs in permeabilised mammalian cells and that the incorporated bromouridine residues do not interfere with pre-rRNA maturation. Our results suggest that the maturation process of ribosomal RNA in permeabilised cells takes place at, or nearby, the site of transcription and that the processing complex is assembled during or early after the rRNA transcription.