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A Multimodal Strategy Used By A Large c-di-GMP Network

Kurt M. Dahlstrom, Alan J. Collins, Georgia Doing, Jacyln N Taroni, Timothy J. Gauvin, Casey S. Greene, Deborah A. Hogan, View ORCID ProfileGeorge A. O’Toole
doi: https://doi.org/10.1101/198481
Kurt M. Dahlstrom
1Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire 03755
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Alan J. Collins
1Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire 03755
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Georgia Doing
1Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire 03755
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Jacyln N Taroni
2Department of Systems Pharmacology and TranslationalTherapeutics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania 19104
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Timothy J. Gauvin
1Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire 03755
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Casey S. Greene
2Department of Systems Pharmacology and TranslationalTherapeutics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania 19104
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Deborah A. Hogan
1Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire 03755
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George A. O’Toole
1Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire 03755
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  • ORCID record for George A. O’Toole
  • For correspondence: georgeo@dartmouth.edu
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Abstract

The Pseudomonas fluorescens genome encodes for 50+ proteins involved in -di-GMP signaling. Here, we demonstrate that when tested across 188 nutrients, these enzymes and effectors appear capable of impacting biofilm formation. Transcriptional analysis of network members across ∼50 nutrient conditions indicates that altered gene expression can explain a subset, but not all, of biofilm-formation responses to the nutrients. Additional organization of the network is likely achieved through physical interaction, as determined via probing ∼2000 interactions by bacterial two-hybrid assays. Our analysis revealed a multimodal regulatory strategy, using combinations of ligand-mediated signals, protein-protein interaction and/or transcriptional regulation to further fine-tune c-di-GMP-mediated responses. These results create a profile of a large c-di-GMP network that is used to make important cellular decisions, opening the door to future model building and the ability to engineer this complex circuitry in other bacteria.

Abstract Importance Cyclic diguanylate (c-di-GMP) is a key signalling molecule regulating bacterial biofilm formation, and many microbes have up to dozens of proteins that make, break or bind this dinucleotide. Thus, a major open question in the field is how signalling specificity is conferred in this context with a soluble signalling molecule. Here, we take a systems approach, using mutational analysis, transcriptional studies and bacterial two-hybrid analysis to interrogate this network. We find that the network typically combines two or more modes of regulation (i.e., transcriptional control with protein-protein interaction) to generate an observed output.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted October 04, 2017.
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A Multimodal Strategy Used By A Large c-di-GMP Network
Kurt M. Dahlstrom, Alan J. Collins, Georgia Doing, Jacyln N Taroni, Timothy J. Gauvin, Casey S. Greene, Deborah A. Hogan, George A. O’Toole
bioRxiv 198481; doi: https://doi.org/10.1101/198481
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A Multimodal Strategy Used By A Large c-di-GMP Network
Kurt M. Dahlstrom, Alan J. Collins, Georgia Doing, Jacyln N Taroni, Timothy J. Gauvin, Casey S. Greene, Deborah A. Hogan, George A. O’Toole
bioRxiv 198481; doi: https://doi.org/10.1101/198481

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