title: Functional Dynamics within the Human Ribosome Regulate the Rate of Active Protein Synthesis.
abstract: The regulation of protein synthesis contributes to gene expression in both normal physiology and disease, yet kinetic investigations of the human translation mechanism are currently lacking. Using single-molecule fluorescence imaging methods, we have quantified the nature and timing of structural processes in human ribosomes during single-turnover and processive translation reactions. These measurements reveal that functional complexes exhibit dynamic behaviors and thermodynamic stabilities distinct from those observed for bacterial systems. Structurally defined sub-states of pre- and post-translocation complexes were sensitive to specific inhibitors of the eukaryotic ribosome, demonstrating the utility of this platform to probe drug mechanism. The application of three-color single-molecule fluorescence resonance energy transfer (smFRET) methods further revealed a long-distance allosteric coupling between distal tRNA binding sites within ribosomes bearing three tRNAs, which contributed to the rate of processive translation.
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA; Tri-Institutional Training Program in Chemical Biology, Weill Cornell Medical College, Rockefeller University, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA.
authors.forename: Angelica
authors.initials: A
authors.lastname: Ferguson
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Leyi
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Roger B
authors.initials: RB
authors.lastname: Altman
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Terry
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Manuel F
authors.initials: MF
authors.lastname: Juette
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Burnett
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Jose L
authors.initials: JL
authors.lastname: Alejo
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Randall A
authors.initials: RA
authors.lastname: Dass
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Parks
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA; Department of Pharmacology and Physiology, Karolinska Institute, 171 77 Stockholm, Sweden.
authors.forename: C Theresa
authors.initials: CT
authors.lastname: Vincent
authors.affiliation: Department of Physiology and Biophysics, Weill Cornell Medical College, New York, NY 10065, USA; Tri-Institutional Training Program in Chemical Biology, Weill Cornell Medical College, Rockefeller University, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA. Electronic address: scb2005@med.cornell.edu.
authors.forename: Scott C
authors.initials: SC
authors.lastname: Blanchard
mesh_terms: D000494:Allosteric Regulation; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D014176:Protein Biosynthesis; D012343:RNA, Transfer / Q000737:chemistry* / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism
chemical_list: D012343:RNA, Transfer