The role of the 3′ external transcribed spacer in yeast pre-rRNA processing

We have undertaken a deletion analysis of the 3' external transcribed spacer (3' ETS) in the pre-rRNA of Saccharomyces cerevisiae. A stem loop structure immediately 3' to the 25 S rRNA region is necessary and sufficient for processing of the 3' ETS. This is believed to be by cotranscriptional cleavage by Rntlp, the yeast homologue of RNase III. Ln addition, this stem-loop is required for cleavage of site A(3) by RNase MRP and for processing at site B-IL in the 3' region of ITS1. Processing at an upstream site in ITS1, site A(2), and at sites in the 5' external transcribed spacer are not affected, even by complete deletion of the 3' ETS. We conclude that processing in the 3' ETS and in ITS1 is coupled. This would constitute a quality control that prevents synthesis of the 5.8 S rRNA and 5' end maturation of the 25 S rRNA in transcripts which are incomplete due to premature transcription termination. (C) 1998 Academic Press Limited.