In vitro sequence-specific cleavage in transcribed spacer of mouse precursor ribosomal RNA

When we compared the sequences near four terminal sites of transcripts including two endoribonucleolytic cleavage sites in external transcribed spacer 1 upstream of the 5' end of the mouse 18S rRNA (Mishima, Y., Mitsuma, T., & Ogata, K. (1985) EMBO J. 4, 3879-3886), a seven-nucleotide consensus sequence, GGPyUUGPy (Py is C or U), was obtained. The results of both in vitro pulse-chase experiments and S1 nuclease mappings using the mouse rDNA fragment of the transcription initiation region indicated that ribonucleolytic cleavages take place in the sequence matching the consensus sequence at more than five nucleotides out of the seven positions. Furthermore, effective ribonucleolytic cleavages in vitro were observed in a sequence, GGCUUGU, in the internal transcribed spacer 2 located between 5.8S and 28S rRNA. These results demonstrate that the ribonucleolytic cleavages occur preferentially in the sequence of GGPyUUGPy in the transcribed spacer regions of the mouse pre-rRNA. From this information, we infer the existence of processing steps for the pre-rRNA maturation involving the sequence-specific ribonucleolytic cleavages.