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Probing Transcription Factor Dynamics at the Single-Molecule Level in a Living Cell
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Probing Transcription Factor Dynamics at the Single-Molecule Level in a Living Cell
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Journal Article
Probing Transcription Factor Dynamics at the Single-Molecule Level in a Living Cell
2007
Overview
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.
Publisher
American Association for the Advancement of Science,The American Association for the Advancement of Science
Subject
/ Bacterial Proteins - metabolism
/ Bacterial Proteins/genetics/metabolism
/ Bacteriological methods and techniques used in bacteriology
/ Biologi
/ Biological and medical sciences
/ Biology
/ Dimers
/ DNA
/ E coli
/ Escherichia coli Proteins - metabolism
/ Fundamental and applied biological sciences. Psychology
/ Kinetics
/ Luminescent Proteins - genetics
/ Luminescent Proteins - metabolism
/ Luminescent Proteins/genetics/metabolism
