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Science 25 May 2007:
Vol. 316. no. 5828, pp. 1191 - 1194
DOI: 10.1126/science.1141967
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Reports

Probing Transcription Factor Dynamics at the Single-Molecule Level in a Living Cell

Johan Elf,1* Gene-Wei Li,2* X. Sunney Xie1{dagger}

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.

1 Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
2 Department of Physics, Harvard University, Cambridge, MA 02138, USA.

* These authors contributed equally to this work.

{dagger} To whom correspondence should be addressed. E-mail: xie{at}chemistry.harvard.edu

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