Translation dynamics of single mRNAs in live cells and neurons.
Translation is the fundamental biological process converting mRNA information into proteins. Single-molecule imaging in live cells has illuminated the dynamics of RNA transcription; however, it is not yet applicable to translation. Here, we report single-molecule imaging of nascent peptides (SINAPS) to assess translation in live cells. The approach provides direct readout of initiation, elongation, and location of translation. We show that mRNAs coding for endoplasmic reticulum (ER) proteins are translated when they encounter the ER membrane. Single-molecule fluorescence recovery after photobleaching provides direct measurement of elongation speed (5 amino acids per second). In primary neurons, mRNAs are translated in proximal dendrites but repressed in distal dendrites and display "bursting" translation. This technology provides a tool with which to address the spatiotemporal translation mechanism of single mRNAs in living cells.
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Bin
B
Wu
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Carolina
C
Eliscovich
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Young J
YJ
Yoon
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Janelia Research Campus, Howard Hughes Medical Institute (HHMI), Ashburn, VA 20147, USA. robert.singer@einstein.yu.edu singerr@janelia.hhmi.org.
Robert H
RH
Singer
D000818:Animals; D045744:Cell Line, Tumor; D003712:Dendrites / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D036681:Fluorescence Recovery After Photobleaching / Q000379:methods*; D006801:Humans; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D009474:Neurons / Q000378:metabolism*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism
D012333:RNA, Messenger