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Probing transcription factor dynamics at the single-molecule level in a living cell.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology.
2007 (English)In: Science, ISSN 0036-8075, E-ISSN 1095-9203, Vol. 316, no 5828, 1191-1194 p.Article in journal (Other academic) Published
Abstract [en]

Transcription factors regulate gene expression through their binding to DNA. In a living Escherichia coli cell, we directly observed specific binding of a lac repressor, labeled with a fluorescent protein, to a chromosomal lac operator. Using single-molecule detection techniques, we measured the kinetics of binding and dissociation of the repressor in response to metabolic signals. Furthermore, we characterized the nonspecific binding to DNA, one-dimensional (1D) diffusion along DNA segments, and 3D translocation among segments through cytoplasm at the single-molecule level. In searching for the operator, a lac repressor spends ~90% of time nonspecifically bound to and diffusing along DNA with a residence time of <5 milliseconds. The methods and findings can be generalized to other nucleic acid binding proteins.

Place, publisher, year, edition, pages
2007. Vol. 316, no 5828, 1191-1194 p.
Keyword [en]
Bacterial Proteins/genetics/*metabolism, DNA; Bacterial/*metabolism, Diffusion, Escherichia coli/*genetics, Escherichia coli Proteins/*metabolism, Kinetics, Lac Operon, Luminescent Proteins/genetics/metabolism, Microscopy; Fluorescence, Operator Regions (Genetics), Protein Binding, Repressor Proteins/*metabolism, Signal Transduction
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:uu:diva-15165DOI: 10.1126/science.1141967PubMedID: 17525339OAI: oai:DiVA.org:uu-15165DiVA: diva2:42936
Available from: 2008-02-06 Created: 2008-02-06 Last updated: 2017-12-11Bibliographically approved

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Publisher's full textPubMedhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Retrieve&list_uids=17525339&dopt=Citation

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Elf, Johan

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