8FTO
E. coli 70S ribosome with an improved MS2 tag inserted in H98
This is a non-PDB format compatible entry.
Summary for 8FTO
| Entry DOI | 10.2210/pdb8fto/pdb |
| EMDB information | 29449 29483 29484 29485 |
| Descriptor | 50S ribosomal protein L33, 30S ribosomal protein S5, 30S ribosomal protein S6, ... (57 entities in total) |
| Functional Keywords | ms2-tag, h98, ribosome |
| Biological source | Escherichia coli More |
| Total number of polymer chains | 54 |
| Total formula weight | 2191354.82 |
| Authors | Nissley, A.J.,Cate, J.H.D. (deposition date: 2023-01-12, release date: 2023-08-02, Last modification date: 2025-02-19) |
| Primary citation | Nissley, A.J.,Kamal, T.S.,Cate, J.H.D. Interactions between terminal ribosomal RNA helices stabilize the E. coli large ribosomal subunit. Rna, 29:1500-1508, 2023 Cited by PubMed Abstract: The ribosome is a large ribonucleoprotein assembly that uses diverse and complex molecular interactions to maintain proper folding. In vivo assembled ribosomes have been isolated using MS2 tags installed in either the 16S or 23S ribosomal RNAs (rRNAs), to enable studies of ribosome structure and function in vitro. RNA tags in the 50S subunit have commonly been inserted into an extended helix H98 in 23S rRNA, as this addition does not affect cellular growth or in vitro ribosome activity. Here, we find that 50S subunits with MS2 tags inserted in H98 are destabilized compared to wild-type (WT) 50S subunits. We identify the loss of RNA-RNA tertiary contacts that bridge helices H1, H94, and H98 as the cause of destabilization. Using cryogenic electron microscopy (cryo-EM), we show that this interaction is disrupted by the addition of the MS2 tag and can be restored through the insertion of a single adenosine in the extended H98 helix. This work establishes ways to improve MS2 tags in the 50S subunit that maintain ribosome stability and investigates a complex RNA tertiary structure that may be important for stability in various bacterial ribosomes. PubMed: 37419664DOI: 10.1261/rna.079690.123 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (1.85 Å) |
Structure validation
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