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Tracking of single tRNAs for translation kinetics measurements in chloramphenicol treated bacteria
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Systems Biology.ORCID iD: 0000-0001-7288-6363
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Systems Biology.ORCID iD: 0000-0003-1591-3702
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Systems Biology.ORCID iD: 0000-0001-8811-2629
2019 (English)In: Methods, ISSN 1046-2023, E-ISSN 1095-9130, Vol. 162-163, p. 23-30Article in journal (Refereed) Published
Abstract [en]

Chloramphenicol is a broad-spectrum antibiotic targeting the protein synthesis machinery by binding to the bacterial ribosome. Chloramphenicol has been considered a classic general inhibitor of translation, blocking the accommodation of aa-tRNA into the A site of the large ribosomal subunit. However, recent studies suggest that this proposed mechanism is a simplification and that the effect of chloramphenicol on mRNA translation is much more dynamic. By tracking single dye-labelled elongator and initiator tRNAs in Escherichia coli cells treated with chloramphenicol, we observe the direct effect of chloramphenicol on translation kinetics. We find clear indications of slow but significant mRNA translation on drug bound ribosomes.

Place, publisher, year, edition, pages
Elsevier, 2019. Vol. 162-163, p. 23-30
Keywords [en]
Protein synthesis, Translation, Single-molecule tracking, tRNA, In vivo, Chloramphenicol, Chloramphenicol resistance
National Category
Biochemistry and Molecular Biology Microbiology
Identifiers
URN: urn:nbn:se:uu:diva-390695DOI: 10.1016/j.ymeth.2019.02.004ISI: 000475837300004PubMedID: 30742999OAI: oai:DiVA.org:uu-390695DiVA, id: diva2:1342532
Funder
Swedish Research Council, 2015-04111Swedish Research Council, 2016-06264Carl Tryggers foundation , 15:243Carl Tryggers foundation , 17:226Available from: 2019-08-13 Created: 2019-08-13 Last updated: 2019-08-13Bibliographically approved

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