A tale of two microbes : nanoscale molecular cinematography of bacteria and coronaviruses
Abstract/Contents
- Abstract
- Since the first optical detection of a single molecule 32 years ago, single-molecule microscopy has revolutionized our understanding of complex chemical and biological systems. Precise localization combined with switchable or blinking fluorescent labels allows super-resolution microscopy to view biological systems down to the ~10 nm scale. In this thesis, I will discuss the hidden secrets of two tiny creatures revealed by single molecule nanoscopic imaging: 1) The bacterium Caulobacter crescentus programs bifunctionality into a regulatory protein to drive asymmetric cell division. 2) Imaging the spatial organization of human coronavirus HCoV-229E genomic RNA and double stranded RNA during infection of lung epithelial cells gives insight into the viral replication process. This thesis will begin with a short introduction on the basic scientific concepts critical to my work and I will go into the theoretical and experimental framework are foundational for research in this dissertation in Chapter 2. Asymmetric cell division yields two distinct daughter cells by mechanisms that underlie cellular diversity in all kingdoms of life. The bacterium Caulobacter crescentus orchestrates this complex process with less than 4000 genes. In Chapter 3, I will describe a strategy deployed by Caulobacter where a regulatory protein, PopA, is programmed to perform distinct roles based on its subcellular address. Combining biochemistry and single-molecule tracking, I will show that depending on the availability of the second messenger molecule, cyclic di-GMP, the PopA protein adopts either a monomer or dimer form. The two oligomeric forms interact with different partners at the two cell poles and mediate the function of two distinct molecular machineries. In addition, I discovered a novel binding partner of PopA at the swarmer pole, which uncovered its additional function. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the viral pathogen causing the ongoing COVID-19 pandemic. In Chapter 4, I will review the development, in collaboration with Stanley Qi lab, of a super-resolution imaging framework that allows us to study coronavirus infection at the nanoscale in a mammalian host cell, using a less infectious model, human coronavirus HCoV-229E. Using this approach, we revealed a striking spatial separation of genomic RNA and double-stranded RNA, which is the intermediate in viral amplification, and showed their distinct structures at different stages of the infection.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2021; ©2021 |
| Publication date | 2021; 2021 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Wang, Jiarui |
|---|---|
| Degree supervisor | Moerner, W. E. (William Esco), 1953- |
| Degree supervisor | Shapiro, Lucy |
| Thesis advisor | Moerner, W. E. (William Esco), 1953- |
| Thesis advisor | Shapiro, Lucy |
| Thesis advisor | Cui, Bianxiao |
| Degree committee member | Cui, Bianxiao |
| Associated with | Stanford University, Department of Chemistry |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Jiarui Wang. |
|---|---|
| Note | Submitted to the Department of Chemistry. |
| Thesis | Thesis Ph.D. Stanford University 2021. |
| Location | https://purl.stanford.edu/nr685sy8586 |
Access conditions
- Copyright
- © 2021 by Jiarui Wang
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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