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Multiple LacI-mediated loops revealed by Bayesian statistics and tethered particle motion
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2014 (English)In: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 42, no 16, 10265-77 p.Article in journal (Refereed) Published
Abstract [en]

The bacterial transcription factor LacI loops DNA by binding to two separate locations on the DNA simultaneously. Despite being one of the best-studied model systems for transcriptional regulation, the number and conformations of loop structures accessible to LacI remain unclear, though the importance of multiple coexisting loops has been implicated in interactions between LacI and other cellular regulators of gene expression. To probe this issue, we have developed a new analysis method for tethered particle motion, a versatile and commonly used in vitro single-molecule technique. Our method, vbTPM, performs variational Bayesian inference in hidden Markov models. It learns the number of distinct states (i.e. DNA-protein conformations) directly from tethered particle motion data with better resolution than existing methods, while easily correcting for common experimental artifacts. Studying short (roughly 100 bp) LacI-mediated loops, we provide evidence for three distinct loop structures, more than previously reported in single-molecule studies. Moreover, our results confirm that changes in LacI conformation and DNA-binding topology both contribute to the repertoire of LacI-mediated loops formed in vitro, and provide qualitatively new input for models of looping and transcriptional regulation. We expect vbTPM to be broadly useful for probing complex protein-nucleic acid interactions.

Place, publisher, year, edition, pages
2014. Vol. 42, no 16, 10265-77 p.
National Category
Biophysics
Research subject
Biophysics
Identifiers
URN: urn:nbn:se:su:diva-108941DOI: 10.1093/nar/gku563ISI: 000344647700012PubMedID: 25120267OAI: oai:DiVA.org:su-108941DiVA: diva2:761548
Funder
Swedish Foundation for Strategic Research Wenner-Gren FoundationsThe Royal Swedish Academy of Sciences
Available from: 2014-11-07 Created: 2014-11-07 Last updated: 2017-12-05Bibliographically approved

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