title: Dynamics of Translation of Single mRNA Molecules In Vivo.
abstract: Regulation of mRNA translation, the process by which ribosomes decode mRNAs into polypeptides, is used to tune cellular protein levels. Currently, methods for observing the complete process of translation from single mRNAs in vivo are unavailable. Here, we report the long-term (>1 hr) imaging of single mRNAs undergoing hundreds of rounds of translation in live cells, enabling quantitative measurements of ribosome initiation, elongation, and stalling. This approach reveals a surprising heterogeneity in the translation of individual mRNAs within the same cell, including rapid and reversible transitions between a translating and non-translating state. Applying this method to the cell-cycle gene Emi1, we find strong overall repression of translation initiation by specific 5' UTR sequences, but individual mRNA molecules in the same cell can exhibit dramatically different translational efficiencies. The ability to observe translation of single mRNA molecules in live cells provides a powerful tool to study translation regulation.
authors.affiliation: Department of Cellular and Molecular Pharmacology, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158-2517, USA.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Yan
authors.affiliation: Hubrecht Institute, The Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center Utrecht, Utrecht 3584CT, the Netherlands.
authors.forename: Tim A
authors.initials: TA
authors.lastname: Hoek
authors.affiliation: Department of Cellular and Molecular Pharmacology, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158-2517, USA.
authors.forename: Ronald D
authors.initials: RD
authors.lastname: Vale
authors.affiliation: Hubrecht Institute, The Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center Utrecht, Utrecht 3584CT, the Netherlands. Electronic address: m.tanenbaum@hubrecht.eu.
authors.forename: Marvin E
authors.initials: ME
authors.lastname: Tanenbaum
mesh_terms: D020121:5' Untranslated Regions; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000378:metabolism; D044783:F-Box Proteins / Q000378:metabolism; D005453:Fluorescence; D017930:Genes, Reporter; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000032:analysis; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis; D061848:Optical Imaging / Q000379:methods*; D010441:Peptide Chain Elongation, Translational; D010442:Peptide Chain Initiation, Translational; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D020121:5' Untranslated Regions; D018797:Cell Cycle Proteins; D044783:F-Box Proteins; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D012333:RNA, Messenger; D000097573:Red Fluorescent Protein; C498960:FBXO5 protein, human