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. 2021:2346:173-182.
doi: 10.1007/7651_2020_306.

Purification of the Vibrio Quorum-Sensing Transcription Factors LuxR, HapR, and SmcR

Jane D Newman  1 , Julia C van Kessel  2
Affiliations

Purification of the Vibrio Quorum-Sensing Transcription Factors LuxR, HapR, and SmcR

Jane D Newman et al. Methods Mol Biol. 2021.

Abstract

Quorum sensing is a cell density-dependent form of cellular communication among bacteria. This signaling process has been heavily studied in vibrios due to their diverse and complex phenotypes and relevance to human and aquaculture disease. Mechanistic studies of Vibrio quorum sensing have required optimization of protein purification techniques to examine the role of key proteins, such as the LuxR/HapR family of transcription factors that control quorum-sensing gene expression. Protein purification is the cornerstone of biochemistry, and it is crucial to consistently produce batches of protein that are pure, active, and concentrated to perform various assays. The methods described here are optimized for purification of the Vibrio master quorum-sensing regulators, LuxR (Vibrio harveyi), HapR (Vibrio cholerae), and SmcR (Vibrio vulnificus). We anticipate that these methods can be applied to other proteins in this family of transcription factors.

Keywords: Affinity purification; HapR; LuxR; Native purification; Protein purification; SmcR; Transcription factor; Vibrio.

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References

    1. Ball AS, Chaparian RR, van Kessel JC (2017) Quorum sensing gene regulation by LuxR/HapR master regulators in vibrios. J Bacteriol 199:JB.00105-17. https://doi.org/10.1128/JB.00105-17 - DOI
    1. Kim Y, Kim BS, Park YJ et al (2010) Crystal structure of SmcR, a quorum-sensing master regulator of Vibrio vulnificus, provides insight into its regulation of transcription. J Biol Chem 285:14020–14030. https://doi.org/10.1074/jbc.M109.100248 - DOI - PubMed - PMC
    1. De Silva RS, Kovacikova G, Lin W et al (2007) Crystal structure of the Vibrio cholerae quorum-sensing regulatory protein HapR. J Bacteriol 189:5683–5691. https://doi.org/10.1128/JB.01807-06 - DOI - PubMed - PMC
    1. Van Kessel JC, Rutherford ST, Shao Y et al (2013) Individual and combined roles of the master regulators apha and luxr in control of the Vibrio harveyi quorum-sensing regulon. J Bacteriol 195:436–443. https://doi.org/10.1128/JB.01998-12 - DOI - PubMed - PMC
    1. Chaparian RR, Olney SG, Hustmyer CM et al (2016) Integration host factor and LuxR synergistically bind DNA to coactivate quorum-sensing genes in Vibrio harveyi. Mol Microbiol 101:823–840. https://doi.org/10.1111/mmi.13425 - DOI - PubMed

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