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The CpxRA Two-Component System is Involved in the Maintenance of the Integrity of the Cell Envelope in the Rumen Bacterium Mannheimia succiniciproducens

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Abstract

In this study, we characterized the CpxRA two-component signal transduction system of the rumen bacterium Mannheimia succiniciproducens. The truncated form of the CpxA sensor kinase protein without its transmembrane domain was able to autophosphorylate and transphosphorylate the CpxR response regulator protein in vitro. We identified 152 putative target genes for the Cpx system in M. succiniciproducens, which were differentially expressed by more than twofold upon overexpression of the CpxR protein. Genes of a putative 16-gene operon related to the cell wall and lipopolysaccharide biosynthesis were induced strongly upon CpxR overexpression. The promoter region of the first gene of this operon, wecC encoding UDP-N-acetyl-D-mannosaminuronate dehydrogenase, was analyzed and found to contain a sequence homologous to the CpxR box of Escherichia coli. An electrophoretic mobility shift assay showed that the phosphorylated CpxR proteins were able to bind specifically to PCR-amplified DNA fragments containing the promoter sequence of wecC. Furthermore, a cpxR-disrupted mutant strain exhibited increased envelope permeability compared with a wild-type strain. These results suggest that the Cpx system of M. succiniciproducens is involved in the maintenance of the integrity of the cell envelope.

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Acknowledgments

This work was supported by Grants (2011-0031947 and 2011-0031963) from the Intelligent Synthetic Biology Global Frontier Program of the National Research Foundation and by a Grant from the Next-Generation BioGreen 21 Program (PJ00952402) of the Rural Development Administration, Republic of Korea.

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Authors and Affiliations

  1. Synthetic Biology and Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea

    Seulgi Yun, Eun-Gyeong Lee, Sang-Yoon Kim, Jong Moon Shin, Won Seok Jung, Doo-Byoung Oh & Ohsuk Kwon

  2. Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon, Republic of Korea

    Seulgi Yun, Doo-Byoung Oh & Ohsuk Kwon

  3. Department of Chemical and Biomolecular Engineering (BK21 Plus Program), Metabolic and Biomolecular Engineering National Research Laboratory, BioProcess Engineering Research Center, Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, KAIST, Daejeon, Republic of Korea

    Sang Yup Lee

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  1. Seulgi Yun
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  2. Eun-Gyeong Lee
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  4. Jong Moon Shin
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Correspondence to Ohsuk Kwon.

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Seulgi Yun and Eun-Gyeong Lee contributed equally to this work.

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Yun, S., Lee, EG., Kim, SY. et al. The CpxRA Two-Component System is Involved in the Maintenance of the Integrity of the Cell Envelope in the Rumen Bacterium Mannheimia succiniciproducens . Curr Microbiol 70, 103–109 (2015). https://doi.org/10.1007/s00284-014-0686-5

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