Applying DNA affinity chromatography to specifically screen for sucrose-related DNA-binding transcriptional regulators of Xanthomonas campestris

Leßmeier L, Alkhateeb R, Schulte F, Steffens T, Loka TP, Pühler A, Niehaus K, Vorhölter F-J (2016)
Journal of Biotechnology 232: 89-98.

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Abstract
At a molecular level, the regulation of many important cellular processes is still obscure in xanthomonads, a bacterial group of outstanding relevance as world-wide plant pathogens and important for biotechnology as producers of the polysaccharide xanthan. Transcriptome analysis indicated a sucrose-dependent regulation of 18 genes in Xanthomonas campestris pv. campestris (Xcc) B100. The expression of 12 of these genes was clearly increased in the presence of sucrose. Only part of these genes was obviously involved in sucrose utilization. To identify regulatory proteins involved in transcriptional regulation, a DNA fragment-specific pull-down approach was established for Xcc. Putative promoter regions were identified and used to isolate DNA-binding proteins, which were separated by SDS PAGE and identified by MALDI-TOF mass spectrometry. This led to the identification of four transcriptional regulators, among them the global transcriptional regulator Clp and a previously identified regulator of sucrose utilization, SuxR, plus a third DNA-binding transcriptional regulator encoded by xcc-b100_2861 and recently shown to interact with a cyclic di-GMP-binding protein. The fourth regulatory protein was encoded by xcc-b100_2791. These results indicate DNA fragment-specific pull-down experiments as promising approaches to screen for specific DNA-binding regulatory proteins in Xcc. (C) 2016 Elsevier B.V. All rights reserved.
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Conference
10th CeBiTec Symposium on Bioinformatics for Biotechnology and Biomedicine
Location
Bielefeld, Germany
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eISSN
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Leßmeier L, Alkhateeb R, Schulte F, et al. Applying DNA affinity chromatography to specifically screen for sucrose-related DNA-binding transcriptional regulators of Xanthomonas campestris. Journal of Biotechnology. 2016;232:89-98.
Leßmeier, L., Alkhateeb, R., Schulte, F., Steffens, T., Loka, T. P., Pühler, A., Niehaus, K., et al. (2016). Applying DNA affinity chromatography to specifically screen for sucrose-related DNA-binding transcriptional regulators of Xanthomonas campestris. Journal of Biotechnology, 232, 89-98. doi:10.1016/j.jbiotec.2016.04.007
Leßmeier, L., Alkhateeb, R., Schulte, F., Steffens, T., Loka, T. P., Pühler, A., Niehaus, K., and Vorhölter, F. - J. (2016). Applying DNA affinity chromatography to specifically screen for sucrose-related DNA-binding transcriptional regulators of Xanthomonas campestris. Journal of Biotechnology 232, 89-98.
Leßmeier, L., et al., 2016. Applying DNA affinity chromatography to specifically screen for sucrose-related DNA-binding transcriptional regulators of Xanthomonas campestris. Journal of Biotechnology, 232, p 89-98.
L. Leßmeier, et al., “Applying DNA affinity chromatography to specifically screen for sucrose-related DNA-binding transcriptional regulators of Xanthomonas campestris”, Journal of Biotechnology, vol. 232, 2016, pp. 89-98.
Leßmeier, L., Alkhateeb, R., Schulte, F., Steffens, T., Loka, T.P., Pühler, A., Niehaus, K., Vorhölter, F.-J.: Applying DNA affinity chromatography to specifically screen for sucrose-related DNA-binding transcriptional regulators of Xanthomonas campestris. Journal of Biotechnology. 232, 89-98 (2016).
Leßmeier, Lennart, Alkhateeb, Rabeaa, Schulte, Fabian, Steffens, Tim, Loka, Tobias Pascal, Pühler, Alfred, Niehaus, Karsten, and Vorhölter, Frank-Jörg. “Applying DNA affinity chromatography to specifically screen for sucrose-related DNA-binding transcriptional regulators of Xanthomonas campestris”. Journal of Biotechnology 232 (2016): 89-98.
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