Interactions of the release factor RF1 with the ribosome as revealed by cryo-EM.
In eubacteria, termination of translation is signaled by any one of the stop codons UAA, UAG, and UGA moving into the ribosomal A site. Two release factors, RF1 and RF2, recognize and bind to the stop codons with different affinities and trigger the hydrolysis of the ester bond that links the polypeptide with the P-site tRNA. Cryo-electron microscopy (cryo-EM) results obtained in this study show that ribosome-bound RF1 is in an open conformation, unlike the closed conformation observed in the crystal structure of the free factor, allowing its simultaneous access to both the decoding center and the peptidyl-transferase center. These results are similar to those obtained for RF2, but there is an important difference in how the factors bind to protein L11, which forms part of the GTPase-associated center of the large ribosomal subunit. The difference in the binding position, C-terminal domain for RF2 versus N-terminal domain for RF1, explains a body of L11 mutation studies that revealed differential effects on the activity of the two factors. Very recent data obtained with small-angle X-ray scattering now reveal that the solution structure of RF1 is open, as here seen on the ribosome by cryo-EM, and not closed, as seen in the crystal.
Rawat
Urmila
U
Howard Hughes Medical Institute, University of Michigan School of Medicine, 1150 West Medical Ctr. Drive, Ann Arbor, MI 48109, USA.
Gao
Haixiao
H
Zavialov
Andrey
A
Gursky
Richard
R
Ehrenberg
Måns
M
Frank
Joachim
J
D020285:Cryoelectron Microscopy; D020558:GTP Phosphohydrolases / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010454:Peptide Termination Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010458:Peptidyl Transferases / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D014176:Protein Biosynthesis; D011487:Protein Conformation; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism; D020124:Thermotoga maritima / Q000378:metabolism
D010454:Peptide Termination Factors; C041460:peptide chain termination release factor 2; D010458:Peptidyl Transferases; D020558:GTP Phosphohydrolases