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Specificity in signaling : bacterial decision-making through the cyclic diguanylate second messenger / by Kurt M. Dahlstrom

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Author	Dahlstrom, Kurt M.
Title	Specificity in signaling : bacterial decision-making through the cyclic diguanylate second messenger / by Kurt M. Dahlstrom
Date	2016
Abstract	Cyclic diguanylate (c-di-GMP) is a primarily bacterial and nearly universal second messenger used to stimulate a variety of responses from the cell by binding effector proteins. It is created by enzymes called diguanylate cyclases (DGCs) and broken down by phosphodiesterases (PDEs). Any given bacterial species that uses this mode of signaling may have one or dozens of these enzymes. Many c-di-GMP-controlled phenotypes have been found to be regulated by specific DGCs or PDEs, and not others. A major question in the field of microbiology has been to uncover how given DGCs and PDEs may specify one bacterial effector or cellular process, but not another. This work presents evidence that a DGC called GcbC from the soil bacterium Pseudomonas fluorescens physical contacts its effector protein LapD in order to promote biofilm formation. This interaction represents the first of a new class of interacting DGCs with c-di-GMP binding effectors. We demonstrate that this specificity of interaction is linked to discrete ɑ-helices found on these proteins, and that these helices can be predictive of interaction in other proteins. We further show that the auto-inhibitory site of GcbC, normally understood to allosterically inhibit cyclase activity, is also a necessary element for interaction of GcbC with LapD, demonstrating for the first time both a positive and negatively regulatory role for this inhibitory site. Physical interaction was further found to be necessary for proper in vivo signaling of GcbC to LapD. Finally, we have analyzed the broader c-di-GMP network by analyzing all the DGCs, PDEs, and effectors in P. fluorescens for physical interaction with each other, and for transcriptional regulation under a variety of conditions. We also tested many of these proteins for their biofilm phenotype under many different carbon conditions to better understand the environmental context within which this system operates to better translate our knowledge from laboratory conditions to an understanding of how these processes occur in nature. Overall, this work represents a significant contribution to our knowledge of bacterial signaling, and offers a path forward to better model how bacteria use large c-di-GMP networks to make decisions about their life choices.
Description	1 online resource (xii, 228 leaves) : illustrations (some color)
Format	application/pdf
College	Geisel School of Medicine at Dartmouth -- Department of Microbiology and Immunology.
Issuing Body	Dartmouth Dissertations are made available through the Dartmouth Digital Library.
Degree	Thesis (Ph.D)--Dartmouth College.
Copyright	Made available by the author under a Creative Commons Attribution-NonCommercial License.; Made available by the Trustees of Dartmouth College under a Creative Commons Attribution-NonCommercial License. https://creativecommons.org/licenses/by-nc/4.0/
DOI	https://doi.org/10.1349/ddlp.1952

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Specificity in signaling : bacterial decision-making through the cyclic diguanylate second messenger / by Kurt M. Dahlstrom

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