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Integration of multiple stimuli-sensing systems to regulate HrpS and type III secretion system in Erwinia amylovora

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Abstract

The bacterial enhancer binding protein (bEBP) HrpS is essential for Erwinia amylovora virulence by activating the type III secretion system (T3SS). However, how the hrpS gene is regulated remains poorly understood in E. amylovora. In this study, 5′ rapid amplification of cDNA ends and promoter deletion analyses showed that the hrpS gene contains two promoters driven by HrpX/HrpY and the Rcs phosphorelay system, respectively. Electrophoretic mobility shift and gene expression assays demonstrated that integration host factor IHF positively regulates hrpS expression through directly binding the hrpX promoter and positively regulating hrpX/hrpY expression. Moreover, hrpX expression was down-regulated in the relA/spoT ((p)ppGpp-deficient) mutant and the dksA mutant, but up-regulated when the wild-type strain was treated with serine hydroxamate, which induced (p)ppGpp-mediated stringent response. Furthermore, the csrA mutant showed significantly reduced transcripts of major hrpS activators, including the hrpX/hrpY, rcsA and rcsB genes, indicating that CsrA is required for full hrpS expression. On the other hand, the csrB mutant exhibited up-regulation of the rcsA and rcsB genes, and hrpS expression was largely diminished in the csrB/rcsB mutant, indicating that the Rcs system is mainly responsible for the increased hrpS expression in the csrB mutant. These findings suggest that E. amylovora recruits multiple stimuli-sensing systems, including HrpX/HrpY, the Rcs phosphorelay system and the Gac–Csr system, to regulate hrpS and T3SS gene expression.

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  1. Department of Crop Sciences, University of Illinois at Urbana-Champaign, 1201 W. Gregory Dr., Urbana, IL, 61801, USA

    Jae Hoon Lee & Youfu Zhao

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  1. Jae Hoon Lee
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  2. Youfu Zhao
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Correspondence to Youfu Zhao.

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This project was supported by the Agriculture and Food Research Initiative Competitive Grants Program Grant no. 2016-67013-24812 from the USDA National Institute of Food and Agriculture.

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Both authors have no conflicts of interest to declare.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Communicated by S. Hohmann.

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Lee, J.H., Zhao, Y. Integration of multiple stimuli-sensing systems to regulate HrpS and type III secretion system in Erwinia amylovora . Mol Genet Genomics 293, 187–196 (2018). https://doi.org/10.1007/s00438-017-1376-3

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