Functional sites of interaction between release factor RF1 and the ribosome.
Translational release factors decipher stop codons in mRNA and activate hydrolysis of peptidyl-tRNA in the ribosome during translation termination. The mechanisms of these fundamental processes are unknown. Here we have mapped the interaction of bacterial release factor RF1 with the ribosome by directed hydroxyl radical probing. These experiments identified conserved domains of RF1 that interact with the decoding site of the 30S ribosomal subunit and the peptidyl transferase site of the 50S ribosomal subunit. RF1 interacts with a binding pocket formed between the ribosomal subunits that is also the interaction surface of elongation factor EF-G and aminoacyl-tRNA bound to the A site. These results provide a basis for understanding the mechanism of stop codon recognition coupled to hydrolysis of peptidyl-tRNA, mediated by a protein release factor.
Wilson
K S
KS
Center for Molecular Biology of RNA, Sinsheimer Laboratories, University of California, Santa Cruz, California 95064, USA. kwilson@darwin.ucsc.edu
Ito
K
K
Noller
H F
HF
Nakamura
Y
Y
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism; D017665:Hydroxyl Radical; D008958:Models, Molecular; D018716:Molecular Mimicry; D015335:Molecular Probes; D010454:Peptide Termination Factors; D010458:Peptidyl Transferases / Q000378:metabolism; D011487:Protein Conformation; D012270:Ribosomes / Q000378:metabolism*; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism*
D001426:Bacterial Proteins; D015335:Molecular Probes; D010454:Peptide Termination Factors; D015534:Trans-Activators; D017665:Hydroxyl Radical; D010458:Peptidyl Transferases; D003545:Cysteine