The 23 S rRNA environment of ribosomal protein L9 in the 50 S ribosomal subunit
| Autor: | K R, Lieberman, M A, Firpo, A J, Herr, T, Nguyenle, J F, Atkins, R F, Gesteland, H F, Noller |
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| Rok vydání: | 2000 |
| Předmět: |
Models
Molecular Ribosomal Proteins Binding Sites Hydroxyl Radical RNA-Binding Proteins Sulfuric Acid Esters Protein Structure Secondary Molecular Weight RNA Bacterial RNA Ribosomal 23S Molecular Probes Mutation Escherichia coli Nucleic Acid Conformation Ferrous Compounds Genetic Engineering Ribosomes Edetic Acid |
| Zdroj: | Journal of molecular biology. 297(5) |
| ISSN: | 0022-2836 |
| Popis: | Ribosomal protein L9 consists of two globular alpha/beta domains separated by a nine-turn alpha-helix. We examined the rRNA environment of L9 by chemical footprinting and directed hydroxyl radical probing. We reconstituted L9, or individual domains of L9, with L9-deficient 50 S subunits, or with deproteinized 23 S rRNA. A footprint was identified in domain V of 23 S rRNA that was mainly attributable to N-domain binding. Fe(II) was tethered to L9 via cysteine residues introduced at positions along the alpha-helix and in the C-domain, and derivatized proteins were reconstituted with L9-deficient subunits. Directed hydroxyl radical probing targeted regions of domains I, III, IV, and V of 23 S rRNA, reinforcing the view that 50 S subunit architecture is typified by interwoven rRNA domains. There was a striking correlation between the cleavage patterns from the Fe(II) probes attached to the alpha-helix and their predicted orientations, constraining both the position and orientation of L9, as well as the arrangement of specific elements of 23 S rRNA, in the 50 S subunit. |
| Databáze: | OpenAIRE |
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