Following the intersubunit conformation of the ribosome during translation in real time.
We report the direct observation of conformational rearrangements of the ribosome during multiple rounds of elongation. Using single-molecule fluorescence resonance energy transfer, we monitored the intersubunit conformation of the ribosome in real time as it proceeds from codon to codon. During each elongation cycle, the ribosome unlocks upon peptide bond formation, then reverts to the locked state upon translocation onto the next codon. Our data reveal both the specific and cumulative effects of antibiotics on individual steps of translation and uncover the processivity of the ribosome as it elongates. Our approach interrogates the precise molecular events occurring at each codon of the mRNA within the full context of ongoing translation.
Aitken
Colin Echeverría
CE
Biophysics Program, Stanford University, Stanford, California, USA.
Puglisi
Joseph D
JD
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001483:Base Sequence; D003062:Codon / Q000737:chemistry / Q000378:metabolism; D004917:Erythromycin / Q000494:pharmacology; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D054681:Ribosome Subunits, Large, Bacterial / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D054680:Ribosome Subunits, Small, Bacterial / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D003062:Codon; D004917:Erythromycin