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- Volkov, IvanUppsala universitet,Molekylär systembiologi (author)
tRNA tracking for direct measurements of protein synthesis kinetics in live cells
- Article/chapterEnglish2018
Publisher, publication year, extent ...
- 2018-05-16
- NATURE PUBLISHING GROUP,2018
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- LIBRIS-ID:oai:DiVA.org:uu-359663
- https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-359663URI
- https://doi.org/10.1038/s41589-018-0063-yDOI
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- Language:English
- Summary in:English
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- Our ability to directly relate results from test-tube biochemical experiments to the kinetics in living cells is very limited. Here we present experimental and analytical tools to directly study the kinetics of fast biochemical reactions in live cells. Dye-labeled molecules are electroporated into bacterial cells and tracked using super-resolved single-molecule microscopy.Trajectories are analyzed by machine-learning algorithms to directly monitor transitions between bound and free states. In particular, we measure the dwell time of tRNAs on ribosomes, and hence achieve direct measurements of translation rates inside living cells at codon resolution. We find elongation rates with tRNA(Phe) that are in perfect agreement with previous indirect estimates, and once fMet-tRNA(fMet) has bound to the 30S ribosomal subunit, initiation of translation is surprisingly fast and does not limit the overall rate of protein synthesis. The experimental and analytical tools for direct kinetics measurements in live cells have applications far beyond bacterial protein synthesis.
Subject headings and genre
- NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
- NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
- MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Cell- och molekylärbiologi hsv//swe
- MEDICAL AND HEALTH SCIENCES Basic Medicine Cell and Molecular Biology hsv//eng
Added entries (persons, corporate bodies, meetings, titles ...)
- Lindén, Martin,1976-Uppsala universitet,Molekylär systembiologi(Swepub:uu)marli467 (author)
- Aguirre, JavierUppsala universitet,Molekylär systembiologi(Swepub:uu)javag704 (author)
- Ieong, Ka-Weng,1985-Uppsala universitet,Molekylärbiologi(Swepub:uu)kawie779 (author)
- Metelev, MikhailUppsala universitet,Molekylär systembiologi(Swepub:uu)mikme204 (author)
- Elf, JohanUppsala universitet,Molekylär systembiologi(Swepub:uu)johaande (author)
- Johansson, MagnusUppsala universitet,Molekylär systembiologi(Swepub:uu)majoh087 (author)
- Uppsala universitetMolekylär systembiologi (creator_code:org_t)
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- In:Nature Chemical Biology: NATURE PUBLISHING GROUP14:6, s. 618-6261552-44501552-4469
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