Abstract
Modulation of signal transduction via binding of the secondary messenger molecule cyclic di-GMP to effector proteins is a near universal regulatory schema in bacteria. In particular, direct binding of c-di-GMP to transcriptional regulators has been shown to alter gene expression of a variety of processes. Here, we illustrate a pull-down-based DNA:protein binding reaction to determine the relative importance of c-di-GMP in the binding affinity of a target protein to specific DNA sequences. Specifically, the pull-down-based assay enables DNA binding to be analyzed with differing concentrations of c-di-GMP in the absence/presence of specific and nonspecific competitors.
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Acknowledgments
This work was supported by a grant from the National Institute of Health (R01AI080710).
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Chambers, J.R., Sauer, K. (2017). Detection of c-di-GMP-Responsive DNA Binding. In: Sauer, K. (eds) c-di-GMP Signaling. Methods in Molecular Biology, vol 1657. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7240-1_23
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DOI: https://doi.org/10.1007/978-1-4939-7240-1_23
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7240-1
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