Understanding and Engineering Chemically Activated Ubiquitin Ligases for High-throughput Detection, Quantification, and Control of Molecules in Yeast
| dc.contributor.author | Chaisupa, Patarasuda | en |
| dc.contributor.committeechair | Wright, Robert Clay | en |
| dc.contributor.committeemember | Senger, Ryan S. | en |
| dc.contributor.committeemember | Hauf, Silke | en |
| dc.contributor.committeemember | Brown, Anne M. | en |
| dc.contributor.committeemember | Bargmann, Bastiaan | en |
| dc.contributor.department | Biological Systems Engineering | en |
| dc.date.accessioned | 2024-06-11T08:00:53Z | en |
| dc.date.available | 2024-06-11T08:00:53Z | en |
| dc.date.issued | 2024-06-10 | en |
| dc.description.abstract | Fungi, diverse and impactful organisms, exert both beneficial and harmful effects on plants, animals, and humans. Certain fungi produce auxin or indole-3-acetic acid (IAA), a crucial plant growth hormone that influences various aspects of plant growth and defense mechanisms. Conversely, pathogenic fungi can produce auxin and manipulate auxin signaling in their host plant to promote fungal virulence and infection progression. Targeting the auxin signaling pathway in pathogenic fungi offers a novel strategy for combating fungal infections in both plants and humans. Nevertheless, the auxin biosynthesis pathway and the role of auxin in fungal symbioses is not fully understood, in part, due to the lack of a tool for measuring intracellular auxin with high spatial and temporal resolution. This dissertation presents the first genetically encoded biosensor engineered from the E3 ubiquitin ligase to detect and quantify intracellular auxin in a Saccharomyces cerevisiae model. The biosensor has been applied to begin studying auxin metabolism and biosynthesis in yeast as well as better understand the plant auxin co-receptor proteins from which it is built. Additionally, the biosensor is re-engineered for application in inducible protein degradation, controlled by auxin. This tool could be applied to identify novel protein targets for disrupting pathogenic fungal species. Overall, this research offers valuable tool and platform for studying auxin biosynthesis pathway, plant protein and auxin signaling as well as intracellular proteins in fungi. | en |
| dc.description.abstractgeneral | Fungi affect plants, animals, and humans, in both beneficial and harmful ways. Some fungi aid other organisms, while others cause illness. Certain fungi produce a hormone called auxin, or indole-3-acetic acid (IAA), which is essential for plant growth and many environmental responses. Auxin can also assist plants in defending against harmful fungi. Conversely, fungi that infect plants can utilize auxin to promote their own growth and spread. Some fungi even produce auxin, possibly aiding in their colonization of plants. In human fungal infection, it is suggested that auxin may be involved in virulent traits and disease progression. Targeting the auxin signaling pathway in harmful fungi presents an innovative approach to combat fungal infections in both plants and humans. However, our understanding of fungal auxin biosynthesis pathways and their role in fungal infections are not fully understood due to the lack of tools to measure auxin in cells efficiently and accurately. This study introduces the first biological tool, called a biosensor, engineered from auxin responsive proteins from plants, to detect and measure intracellular auxin in Baker's yeast. The biosensor has been used to investigate auxin production by yeast. Additionally, the biosensor has been re-engineered for application in inducible protein degradation, controlled by auxin. This tool could be applied to identify novel protein targets for disrupting pathogenic fungal species. Overall, this research provides useful tool and platform to study auxin production, plant protein function and particular proteins in fungi. | en |
| dc.description.degree | Doctor of Philosophy | en |
| dc.format.medium | ETD | en |
| dc.identifier.other | vt_gsexam:40606 | en |
| dc.identifier.uri | https://hdl.handle.net/10919/119380 | en |
| dc.language.iso | en | en |
| dc.publisher | Virginia Tech | en |
| dc.rights | Creative Commons Attribution 4.0 International | en |
| dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | en |
| dc.subject | biosensors | en |
| dc.subject | F-box protein | en |
| dc.subject | auxin | en |
| dc.subject | protein degradation | en |
| dc.subject | toolkit | en |
| dc.subject | metabolic engineering | en |
| dc.subject | synthetic biology | en |
| dc.title | Understanding and Engineering Chemically Activated Ubiquitin Ligases for High-throughput Detection, Quantification, and Control of Molecules in Yeast | en |
| dc.type | Dissertation | en |
| thesis.degree.discipline | Biological Systems Engineering | en |
| thesis.degree.grantor | Virginia Polytechnic Institute and State University | en |
| thesis.degree.level | doctoral | en |
| thesis.degree.name | Doctor of Philosophy | en |