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Control of type III protein secretion using a minimal genetic system

Author

Listed:
  • Miryoung Song

    (Massachusetts Institute of Technology, Synthetic Biology Center)

  • David J. Sukovich

    (Massachusetts Institute of Technology, Synthetic Biology Center)

  • Luciano Ciccarelli

    (Center for Structural Systems Biology (CSSB), University Medical Center Hamburg-Eppendorf (UKE)
    Deutsches Elektronen-Synchrotron Zentrum (DESY)
    Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna Biocenter (VBC)
    Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC))

  • Julia Mayr

    (Center for Structural Systems Biology (CSSB), University Medical Center Hamburg-Eppendorf (UKE)
    Deutsches Elektronen-Synchrotron Zentrum (DESY)
    Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna Biocenter (VBC)
    Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC))

  • Jesus Fernandez-Rodriguez

    (Massachusetts Institute of Technology, Synthetic Biology Center)

  • Ethan A. Mirsky

    (Massachusetts Institute of Technology, Synthetic Biology Center)

  • Alex C. Tucker

    (Massachusetts Institute of Technology, Synthetic Biology Center)

  • D. Benjamin Gordon

    (MIT-Broad Foundry, Broad Institute of MIT and Harvard)

  • Thomas C. Marlovits

    (Center for Structural Systems Biology (CSSB), University Medical Center Hamburg-Eppendorf (UKE)
    Deutsches Elektronen-Synchrotron Zentrum (DESY)
    Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna Biocenter (VBC)
    Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC))

  • Christopher A. Voigt

    (Massachusetts Institute of Technology, Synthetic Biology Center)

Abstract

Gram-negative bacteria secrete proteins using a type III secretion system (T3SS), which functions as a needle-like molecular machine. The many proteins involved in T3SS construction are tightly regulated due to its role in pathogenesis and motility. Here, starting with the 35 kb Salmonella pathogenicity island 1 (SPI-1), we eliminated internal regulation and simplified the genetics by removing or recoding genes, scrambling gene order and replacing all non-coding DNA with synthetic genetic parts. This process results in a 16 kb cluster that shares no sequence identity, regulation or organizational principles with SPI-1. Building this simplified system led to the discovery of essential roles for an internal start site (SpaO) and small RNA (InvR). Further, it can be controlled using synthetic regulatory circuits, including under SPI-1 repressing conditions. This work reveals an incredible post-transcriptional robustness in T3SS assembly and aids its control as a tool in biotechnology.

Suggested Citation

  • Miryoung Song & David J. Sukovich & Luciano Ciccarelli & Julia Mayr & Jesus Fernandez-Rodriguez & Ethan A. Mirsky & Alex C. Tucker & D. Benjamin Gordon & Thomas C. Marlovits & Christopher A. Voigt, 2017. "Control of type III protein secretion using a minimal genetic system," Nature Communications, Nature, vol. 8(1), pages 1-9, August.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_ncomms14737
    DOI: 10.1038/ncomms14737
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