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The structure of the ITS2-proximal stem is required for pre-rRNA processing in yeast
1National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases, Genetics and Biochemistry Branch, Bethesda, Maryland 20892, USA. bp51h@nih.gov
RNA (New York, N.Y.)
|December 16, 1998
Summary
Formation of the ITS2-proximal stem is essential for efficient pre-ribosomal RNA (rRNA) processing in yeast. Mutations disrupting this stem inhibit maturation, highlighting its conserved role in ribosome biogenesis.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Accurate processing of precursor ribosomal RNA (pre-rRNA) is vital for cell growth, requiring both trans-acting factors and cis-acting RNA elements.
- The ITS2-proximal stem, formed by interactions between 5.8S rRNA and 25S rRNA sequences, is a critical cis-acting element in pre-rRNA processing.
Purpose of the Study:
- To investigate the functional requirement of the ITS2-proximal stem in yeast pre-rRNA processing.
- To determine if mutations affecting ITS2-proximal stem formation impact ribosome biogenesis.
Main Methods:
- Directed mutagenesis of yeast pre-rRNA genes to create mutations affecting the ITS2-proximal stem.
- Expression of mutated pre-rRNAs in yeast.
- Analysis of pre-rRNA processing intermediates and mature rRNA levels using Northern blotting or similar techniques.
Main Results:
- Mutations preventing ITS2-proximal stem formation significantly inhibited pre-rRNA processing.
- Compensatory mutations restoring stem structure partially restored processing efficiency.
- Reduced processing led to decreased mature 5.8S and 25S rRNA and accumulation of 35S pre-rRNA and intermediates.
Conclusions:
- The ITS2-proximal stem is a crucial cis-acting element for efficient yeast pre-rRNA processing.
- The observed phenotype resembles that of U8 snoRNA depletion in vertebrates, suggesting a conserved role for this stem in ribosome biogenesis across species.
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