Whitlock, Sara
(2021)
Designing Synthetic Biomolecular Condensates in Bacteria.
Master's Thesis, University of Pittsburgh.
(Unpublished)
Abstract
Despite lacking many of the organelles possessed by eukaryotes, bacteria are able to localize important enzymes and signaling proteins to specific subcellular locations. One way that bacteria accomplish this localization is through the formation of phase separated membraneless organelles called biomolecular condensates. These structures are composed of proteins and RNA and they self-assemble into multimeric networks, bind client proteins, and recruit them to particular locations within the cell. This thesis describes the history of biomolecular condensates and the design of synthetic mimics of bacterial biomolecular condensates.
Chapter one details the proposed connection between biomolecular condensates and the earliest forms of life on earth. I trace the history of biomolecular condensates starting with the 1924 proposal that complex coacervates—similar to modern biomolecular condensates—were the first forms of life. I then follow the modernization of this idea after decades of ridicule, describing the first discovery, in C. elegans, of a coacervate-like structure operating within the cells of a modern organism. This discovery led to the characterization of biomolecular condensate structures throughout the eukaryotic cell and, in 2018, to the discovery of a biomolecular condensate in bacteria.
With several bacterial condensates now described, I propose in chapter two a strategy for designing synthetic biomolecular condensates in bacteria. These designs are inspired by the pole organizing protein Z (PopZ) of Caulobacter crescentus, which forms a unipolar biomolecular condensate when expressed in Escherichia coli, and by a series of peptide hydrogels designed for cell encapsulation and drug delivery. I use these synthetic constructs to define the minimal domains necessary to form a pole-localized scaffold and demonstrate the impact that varying charge, amount of disorder, and degree of multivalency has on synthetic condensate formation. I recruit and exclude client proteins from synthetic condensates, and I propose ways to use these designs to set up asymmetric division of biomolecular condensates in E. coli.
Overall, this thesis contributes to the understanding of how bacterial biomolecular condensates localize to the pole and develops strategies to mimic this behavior with synthetic condensates.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
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| ETD Committee: |
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| Date: |
20 January 2021 |
| Date Type: |
Publication |
| Defense Date: |
25 November 2020 |
| Approval Date: |
20 January 2021 |
| Submission Date: |
3 December 2020 |
| Access Restriction: |
2 year -- Restrict access to University of Pittsburgh for a period of 2 years. |
| Number of Pages: |
90 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
Dietrich School of Arts and Sciences > Molecular Biophysics and Structural Biology |
| Degree: |
MS - Master of Science |
| Thesis Type: |
Master's Thesis |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
Biomolecular Condensates, Bacteria, E coli, C crescentus, Membraneless Organelle, Synthetic Biology |
| Date Deposited: |
20 Jan 2021 19:41 |
| Last Modified: |
20 Jan 2023 06:15 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/39982 |
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