RNAP II produces capped 18S and 25S ribosomal RNAs resistant to 5′-monophosphate dependent processive 5′ to 3′ exonuclease in polymerase switched Saccharomyces cerevisiae

Abstract

Background: We have previously found that, in the pathogenic yeast Candida albicans, 18S and 25S ribosomal RNA components, containing more than one phosphate on their 5′-end were resistant to 5′-monophosphate requiring 5′ → 3″ exonuclease. Several lines of evidence pointed to RNAP II as the enzyme producing them. Results: We now show the production of such 18S and 25S rRNAs in Saccharomyces cerevisiae that have been permanently switched to RNAP II (due to deletion of part of RNAP I upstream activator alone, or in combination with deletion of one component of RNAP I itself). They contain more than one phosphate at their 5′-end and an anti-cap specific antibody binds to them indicating capping of these molecules. These molecules are found in RNA isolated from nuclei, therefore are unlikely to have been modified in the cytoplasm. Conclusions: Our data confirm the existence of such molecules and firmly establish RNAP II playing a role in their production. The fact that we see these molecules in wild type Saccharomyces cerevisiae indicates that they are not only a result of mutations but are part of the cells physiology. This adds another way RNAP II is involved in ribosome production in addition to their role in the production of ribosome associated proteins.