eprintid: 40086 rev_number: 12 userid: 10137 dir: disk0/00/04/00/86 datestamp: 2021-01-20 18:33:29 lastmod: 2021-01-20 18:33:29 status_changed: 2021-01-20 18:33:29 type: thesis_degree succeeds: 39949 metadata_visibility: show contact_email: duvallsam1@gmail.com item_issues_id: thesis_degree_versioning item_issues_id: duplicate_title_39868 item_issues_id: duplicate_title_39948 item_issues_id: duplicate_title_39949 item_issues_type: thesis_degree_versioning item_issues_type: duplicate_title item_issues_type: duplicate_title item_issues_type: duplicate_title item_issues_description: ETD 40086 is using versioning. item_issues_description: Duplicate title to Duvall, Samuel Development of methods to study bacterial cell signaling in vivo. Doctoral Dissertation, University of Pittsburgh. (Unpublished) item_issues_description: Duplicate title to Duvall, Samuel Development of methods to study bacterial cell signaling in vivo. Doctoral Dissertation, University of Pittsburgh. (Unpublished) item_issues_description: Duplicate title to Duvall, Samuel Development of methods to study bacterial cell signaling in vivo. Doctoral Dissertation, University of Pittsburgh. (Unpublished) item_issues_timestamp: 2020-11-11 07:02:23 item_issues_timestamp: 2020-11-30 07:02:25 item_issues_timestamp: 2020-11-30 07:02:25 item_issues_timestamp: 2020-12-13 07:02:25 item_issues_status: discovered item_issues_status: autoresolved item_issues_status: autoresolved item_issues_status: autoresolved item_issues_count: 1 eprint_status: archive creators_name: Duvall, Samuel creators_email: swd11@pitt.edu creators_id: swd11 title: Development of methods to study bacterial cell signaling in vivo ispublished: unpub divisions: sch_as_chemistry full_text_status: restricted keywords: N/A 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. date: 2021-01-20 date_type: published pages: 349 institution: University of Pittsburgh refereed: TRUE etdcommittee_type: committee_chair etdcommittee_type: committee_member etdcommittee_type: committee_member etdcommittee_type: committee_member etdcommittee_name: Childers, Seth etdcommittee_name: Saxena, Sunil etdcommittee_name: Weber, Steve etdcommittee_name: McCormick, Joseph etdcommittee_email: wschild@pitt.edu etdcommittee_email: sksaxena@pitt.edu etdcommittee_email: sweber@pitt.edu etdcommittee_email: mccormick@duq.edu etdcommittee_id: wschild etdcommittee_id: sksaxena etdcommittee_id: sweber etd_defense_date: 2020-11-23 etd_approval_date: 2021-01-20 etd_submission_date: 2020-10-26 etd_release_date: 2021-01-20 etd_access_restriction: 2_year etd_patent_pending: FALSE thesis_type: dissertation degree: PhD citation: Duvall, Samuel (2021) Development of methods to study bacterial cell signaling in vivo. Doctoral Dissertation, University of Pittsburgh. (Unpublished) document_url: http://d-scholarship-dev.library.pitt.edu/40086/45/Samuel%20Duvall%20Final%20ETD.pdf