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Development of methods to study bacterial cell signaling in vivo

Duvall, Samuel (2021) Development of methods to study bacterial cell signaling in vivo. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Abstract

Our understanding of bacterial signaling systems is still in its early stages. As we learn more about how bacteria respond to environmental and intracellular cues, we will be able to tackle some of the most important problems related to bacteria. Drug-resistance, agriculture, environmental remediation, and more global problems will be better understood and potentially solved in some way by our understanding of bacterial signaling systems. Two-component systems are the foundation of bacterial signaling. Currently, we lack tools for studying two-component signaling systems in vivo and with greater depth than simply whether two proteins colocalize or not. In this dissertation, I will explore the development of tools to study members of two-component signaling systems and the proteins that scaffold them.
Chapter 2 will explore the first example of a histidine kinase FRET sensor which showcases the potential for this type of FRET sensor for any histidine kinase. I provide details on its design and controls for working with it on conventional epifluorescent microscopes. Chapter 3 expands on the use of the CckA-FRET biosensor and previously reported leucine zipper technology to understand the domain interactions of the pseudokinase DivL with the histidine kinase CckA. In Chapter 4, I apply knowledge of histidine kinases to the current antibiotic crisis.
Scaffolding proteins play a multitude of roles in bacteria. In Chapter 5 I will explore PodJ, the multirole scaffolding protein that can recruit PopZ in order to add depth to a signaling network. Chapter 6 will further look at an interesting phenotype of PodJ and how it relates to self-assembly and curvature recognition. Chapter 6 will also cover new tool development for working in C. crescentus.
In each chapter, I will cover questions and future aims. This dissertation overall is aimed at the development of new ways of thinking about bacterial signaling systems and how to study them.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Duvall, Samuelswd11@pitt.eduswd11
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairChilders, Sethwschild@pitt.eduwschild
Committee MemberSaxena, Sunilsksaxena@pitt.edusksaxena
Committee MemberWeber, Stevesweber@pitt.edusweber
Committee MemberMcCormick, Josephmccormick@duq.edu
Date: 20 January 2021
Date Type: Publication
Defense Date: 23 November 2020
Approval Date: 20 January 2021
Submission Date: 26 October 2020
Access Restriction: 2 year -- Restrict access to University of Pittsburgh for a period of 2 years.
Number of Pages: 349
Institution: University of Pittsburgh
Schools and Programs: Dietrich School of Arts and Sciences > Chemistry
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: N/A
Date Deposited: 20 Jan 2021 18:33
Last Modified: 20 Jan 2021 18:33
URI: http://d-scholarship.pitt.edu/id/eprint/40086

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