Conformation of 4.5S RNA in the signal recognition particle and on the 30S ribosomal subunit
| Autor: | Jens Warnecke, Marina V. Rodnina, Johannes Jöckel, Yuri P. Semenkov, Philipp Beinker, Shan-Qing Gu, Wolfgang Wintermeyer |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2005 |
| Předmět: |
Ribosomal Proteins
Molecular Sequence Data Calorimetry Biology Ribosome Article 5S ribosomal RNA RNA Transfer Escherichia coli Signal recognition particle RNA Molecular Biology 50S Base Sequence Eukaryotic Large Ribosomal Subunit Escherichia coli Proteins RNA Molecular biology Protein Structure Tertiary Protein Subunits RNA Bacterial RNA Ribosomal Transfer RNA Biophysics Nucleic Acid Conformation Eukaryotic Ribosome Ribosomes Signal Recognition Particle |
| Zdroj: | RNA |
| Popis: | The signal recognition particle (SRP) from Escherichia coli consists of 4.5S RNA and protein Ffh. It is essential for targeting ribosomes that are translating integral membrane proteins to the translocation pore in the plasma membrane. Independently of Ffh, 4.5S RNA also interacts with elongation factor G (EF-G) and the 30S ribosomal subunit. Here we use a cross-linking approach to probe the conformation of 4.5S RNA in SRP and in the complex with the 30S ribosomal subunit and to map the binding site. The UV-activatable cross-linker p-azidophenacyl bromide (AzP) was attached to positions 1, 21, and 54 of wild-type or modified 4.5S RNA. In SRP, cross-links to Ffh were formed from AzP in all three positions in 4.5S RNA, indicating a strongly bent conformation in which the 5′ end (position 1) and the tetraloop region (including position 54) of the molecule are close to one another and to Ffh. In ribosomal complexes of 4.5S RNA, AzP in both positions 1 and 54 formed cross-links to the 30S ribosomal subunit, independently of the presence of Ffh. The major cross-linking target on the ribosome was protein S7; minor cross-links were formed to S2, S18, and S21. There were no cross-links from 4.5S RNA to the 50S subunit, where the primary binding site of SRP is located close to the peptide exit. The functional role of 4.5S RNA binding to the 30S subunit is unclear, as the RNA had no effect on translation or tRNA translocation on the ribosome. |
| Databáze: | OpenAIRE |
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