Graduate Thesis Or Dissertation
Regulation of AP-3 Vesicle Trafficking by Intrinsically Disordered Hinge Regions Public Deposited
- Abstract
Vesicles bud from maturing Golgi cisternae in a programmed sequence. Budding is mediated by adaptors that recruit cargoes and facilitate vesicle biogenesis. In Saccharomyces cerevisiae, the AP-3 adaptor complex directs cargo from the Golgi to the lysosomal vacuole. The AP-3 core consists of small and medium subunits complexed with two non-identical large subunits, β3 and δ. The C termini of β3 and δ are thought to be flexible hinges linking the core to ear domains that bind accessory proteins involved in vesicular transport. I predicted by computational modeling that the yeast β3 and δ hinges are intrinsically disordered in their entirety and lack ear domains. When either hinge is truncated, AP-3 is recruited to Golgi, but vesicle budding is impaired, and cargoes normally sorted into the AP-3 pathway are mistargeted. This budding deficiency causes AP-3 to accumulate on ring-like Golgi structures with GGA adaptors, which normally function downstream of AP-3 during Golgi maturation. Thus, both disordered hinges of yeast AP-3 have a crucial role in mediating the formation of transport vesicles that bud from the Golgi. Combined analyses of proximal labeling, AP-3 hinge binding assays, and AP-3 cargo sorting identified more than 30 potential regulators of AP-3 trafficking. Among these, the septin proteins encoded by CDC genes were characterized as binding to AP-3 hinges and being required for AP-3 trafficking. Together, these data indicate disordered AP-3 hinge regions perform essential roles in binding accessory proteins required for AP-3 vesicle trafficking despite their lack of ear domains.
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- 2024-07-18
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- 2024-12-18
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Leih_colorado_0051E_19009.pdf | 2024-12-13 | Public | Download |
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Thesis_Approval_Form.pdf | 2024-12-13 | Public | Download |
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Supplemental_Movie_2_1.mp4 | 2024-12-13 | Public | Download |
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Supplemental_Movie_2_2.mp4 | 2024-12-13 | Public | Download |
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Supplemental_Movie_2_4.mp4 | 2024-12-13 | Public | Download |