The cAMP receptor-like protein CLP is a novel c-di-GMP receptor linking cell-cell signaling to virulence gene expression in Xanthomonas campestris

Ko-Hsin Chin, Yen-Chung Lee, Zhi-Le Tu, Chih-Hua Chen, Yi-Hsiung Tseng, Jinn-Moon Yang, Robert P. Ryan, Yvonne McCarthy, John Maxwell Dow, Andrew H.-J. Wang, Shan-Ho Chou

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    127 Citations (Scopus)

    Abstract

    Cyclic-di-GMP [bis-(3'-5')-cyclic diguanosine monophosphate] controls a wide range of functions in eubacteria, yet little is known about the underlying regulatory mechanisms. In the plant pathogen Xanthomonas campestris, expression of a subset of virulence genes is regulated by c-di-GMP and also by the CAP (catabolite activation protein)-like protein XcCLP, a global regulator in the CRP/FNR superfamily. Here, we report structural and functional insights into the interplay between XcCLP and c-di-GMP in regulation of gene expression. XcCLP bound target promoter DNA with submicromolar affinity in the absence of any ligand. This DNAbinding capability was abrogated by c-di-GMP, which bound to XcCLP with micromolar affinity. The crystal structure of XcCLP showed that the protein adopted an intrinsically active conformation for DNA binding.  Alteration of residues of XcCLP implicated in c-di-GMP binding through modeling studies caused a substantial reduction in binding affinity for the nucleotide and rendered DNA binding by these variant proteins insensitive to inhibition by c-di-GMP. Together, these findings reveal the structural mechanism behind a novel class of c-di-GMP effector proteins in the CRP/FNR superfamily and indicate that XcCLP regulates bacterial virulence gene expression in a manner negatively controlled by the c-di-GMP concentrations.

    Original languageEnglish
    Pages (from-to)646-662
    Number of pages17
    JournalJournal of Molecular Biology
    Volume396
    Issue number3
    DOIs
    Publication statusPublished - 26 Feb 2010

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  • Cite this

    Chin, K-H., Lee, Y-C., Tu, Z-L., Chen, C-H., Tseng, Y-H., Yang, J-M., Ryan, R. P., McCarthy, Y., Dow, J. M., Wang, A. H-J., & Chou, S-H. (2010). The cAMP receptor-like protein CLP is a novel c-di-GMP receptor linking cell-cell signaling to virulence gene expression in Xanthomonas campestris. Journal of Molecular Biology, 396(3), 646-662. https://doi.org/10.1016/j.jmb.2009.11.076