Defining the inside and outside of a Catalytic RNA Molecule

• Published in: Science (American Association for the Advancement of Science) Vol. 245; no. 4915; pp. 276 - 282
• Main Authors: Latham, John A., Cech, Thomas R.
• Format: Journal Article
• Language: English
• Published: Washington, DC The American Association for the Advancement of Science 21.07.1989; American Association for the Advancement of Science
• Subjects: Animals; Autoradiography; Base Sequence; Binding Sites; Biochemistry; Biochemistry. Physiology. Immunology. Molecular biology; Biological and medical sciences; Catalytic RNA; Crystallography; Edetic Acid; Electrophoresis, Polyacrylamide Gel; Enzymes; Ferrous Compounds; Fundamental and applied biological sciences. Psychology; Genetics; Molecular Sequence Data; Molecular Structure; Molecules; Nucleic Acid Conformation; Nucleic acids; Nucleotides; Protozoa; Reagents; RNA; RNA Splicing; RNA, Catalytic; RNA, Fungal - analysis; RNA, Ribosomal - analysis; RNA, Ribosomal - metabolism; RNA, Transfer, Phe - analysis; Rock cleavage; Tetrahymena; Tetrahymena - genetics; Transfer RNA; Protozoa; Molecular hybridization; Tetrahymena; Radiolabelling; RNA; Gel electrophoresis; Enzyme; Ciliata; Spatial structure
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.2501870

Ribozymes are RNA molecules that catalyze biochemical reactions. Fe(II)-EDTA, a solvent-based reagent which cleaves both double- and single-stranded RNA, was used to investigate the structure of the Tetrahymena ribozyme. Regions of cleavage alternate with regions of substantial protection along the entire RNA molecule. In particular, most of the catalytic core shows greatly reduced cleavage. These data constitute experimental evidence that an RNA enzyme, like a protein enzyme, has an interior and an exterior. Determination of positions where the phosphodiester backbone of the RNA is on the inside or on the outside of the molecule provides major constraints for modeling the three-dimensional structure of the Tetrahymena ribozyme. This approach should be generally informative for structured RNA molecules.