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TitleExtensive ribosome and RF2 rearrangements during translation termination.
Journal, issue, pagesElife, Vol. 8, Year 2019
Publish dateSep 12, 2019
AuthorsEgor Svidritskiy / Gabriel Demo / Anna B Loveland / Chen Xu / Andrei A Korostelev /
PubMed AbstractProtein synthesis ends when a ribosome reaches an mRNA stop codon. Release factors (RFs) decode the stop codon, hydrolyze peptidyl-tRNA to release the nascent protein, and then dissociate to allow ...Protein synthesis ends when a ribosome reaches an mRNA stop codon. Release factors (RFs) decode the stop codon, hydrolyze peptidyl-tRNA to release the nascent protein, and then dissociate to allow ribosome recycling. To visualize termination by RF2, we resolved a cryo-EM ensemble of 70S•RF2 structures at up to 3.3 Å in a single sample. Five structures suggest a highly dynamic termination pathway. Upon peptidyl-tRNA hydrolysis, the CCA end of deacyl-tRNA departs from the peptidyl transferase center. The catalytic GGQ loop of RF2 is rearranged into a long β-hairpin that plugs the peptide tunnel, biasing a nascent protein toward the ribosome exit. Ribosomal intersubunit rotation destabilizes the catalytic RF2 domain on the 50S subunit and disassembles the central intersubunit bridge B2a, resulting in RF2 departure. Our structures visualize how local rearrangements and spontaneous inter-subunit rotation poise the newly-made protein and RF2 to dissociate in preparation for ribosome recycling.
External linksElife / PubMed:31513010 / PubMed Central
MethodsEM (single particle)
Resolution3.3 - 4.2 Å
Structure data

EMDB-20048, PDB-6ofx:
Non-rotated ribosome (Structure I)
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-20052, PDB-6og7:
70S termination complex with RF2 bound to the UGA codon. Non-rotated ribosome with RF2 bound (Structure II)
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-20056, PDB-6ogf:
70S termination complex with RF2 bound to the UGA codon. Partially rotated ribosome with RF2 bound (Structure III).
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-20057, PDB-6ogg:
70S termination complex with RF2 bound to the UGA codon. Rotated ribosome with RF2 bound (Structure IV).
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-20058, PDB-6ogi:
70S termination complex with RF2 bound to the UAG codon. Rotated ribosome conformation (Structure V)
Method: EM (single particle) / Resolution: 3.4 Å

Chemicals

ChemComp-FME:
N-FORMYLMETHIONINE / N-Formylmethionine

Source
  • escherichia coli (strain k12) (bacteria)
  • escherichia coli (E. coli)
  • escherichia coli k-12 (bacteria)
  • synthetic construct (others)
KeywordsRIBOSOME / Ribosome recycling / translation termination / RF2 / intersubunit rotation

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