Structure of the E. coli ribosome-EF-Tu complex at < 3 angstrom resolution by C-s-corrected cryo-EM

作者：Fischer, Niels^*; Neumann, Piotr; Konevega, Andrey L.; Bock, Lars V.; Ficner, Ralf; Rodnina, Marina V.; Stark, Holger

来源：Nature, 2015, 520(7548): 567-+.

DOI：10.1038/nature14275

• PEPTIDYL TRANSFERASE CENTER
• AMINOACYL-TRANSFER-RNA
• ELONGATION-FACTOR TU
• CRYSTAL-STRUCTURE
• TRANSLATION
• METHYLATIONS
• REFINEMENT
• RESISTANCE
• INITIATION
• MECHANISM

Single particle electron cryomicroscopy (cryo-EM) has recently made significant progress in high-resolution structure determination of macromolecular complexes due to improvements in electron microscopic instrumentation and computational image analysis. However, cryo-EM structures can be highly non-uniform in local resolution" and all structures available to date have been limited to resolutions above 3 angstrom(3,4). Here we present the cryo-EM structure of the 70S ribosome from Escherichia coli in complex with elongation factor Tu, aminoacyl-tRNA and the antibiotic kirromycin at 2.65-2.9 angstrom resolution using spherical aberration (c)-corrected cryo-EM. Overall, the cryo-EM reconstruction at 2.9 angstrom resolution is comparable to the best-resolved X-ray structure of the E. coil 70S ribosome(5) (2.8 angstrom), but provides more detailed information (2.65 angstrom) at the functionally important ribosomal core. The cryo-EM map elucidates for the first time the structure of all 35 rRNA modifications in the bacterial ribosome, explaining their roles in fine-tuning ribosome structure and function and modulating the action of antibiotics. We also obtained atomic models for flexible parts of the ribosome such as ribosomal proteins L9 and L31. The refined cryo-EM -based model presents the currently most complete high-resolution structure of the E. coil ribosome, which demonstrates the power of cryo-EM in structure determination of large and dynamic macromolecular complexes.

• 出版日期2015-4-23