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Folding of circularly permuted transfer RNAs
T Pan1, R R Gutell, O C Uhlenbeck
1Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309.
Science (New York, N.Y.)
|November 29, 1991
Summary
Researchers identified all cleavable ribose-phosphate linkages in yeast transfer RNA (tRNA) without impacting molecular folding. This study reveals the tRNA folding motif can appear internally in other RNA sequences.
Area of Science:
- Molecular Biology
- RNA Structure and Function
- Biochemistry
Background:
- Transfer RNA (tRNA) molecules possess complex three-dimensional structures essential for their biological functions.
- Understanding the specific linkages critical for maintaining tRNA folding is crucial for deciphering RNA structure-function relationships.
Purpose of the Study:
- To identify all ribose-phosphate linkages in yeast tRNA(Phe) that can be cleaved without disrupting the molecule's overall fold.
- To investigate the prevalence and potential internal occurrence of the tRNA folding motif in other RNA sequences.
Main Methods:
- Circular permutation analysis was employed on yeast tRNA(Phe).
- Molecules underwent limited alkaline hydrolysis to introduce a single random break.
- Correctly folded tRNA was identified using lead cleavage at neutral pH, a known indicator of proper tRNA(Phe) folding.
Main Results:
- A comprehensive map of cleavable ribose-phosphate linkages was generated in a single experiment.
- A surprising majority of the circularly permuted tRNA molecules maintained correct folding.
- A subsequent computer search identified numerous instances of similar folding motifs within other RNA sequences in Genbank.
Conclusions:
- The study successfully mapped labile linkages in yeast tRNA(Phe), providing insights into its structural integrity.
- The findings suggest that the characteristic tRNA folding motif can be found as an internal structural element in diverse RNA molecules.
- This discovery opens new avenues for understanding RNA folding principles beyond canonical tRNA structures.
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