Date of Award

3-2023

Document Type

Dissertation

Degree Name

Ph.D.

Organizational Unit

College of Natural Science and Mathematics, Biological Sciences

First Advisor

Erich Kushner

Second Advisor

J. Todd Blankenship

Third Advisor

Martin Margittai

Fourth Advisor

Joe Angleson

Keywords

Intracellular protein trafficking, GTPase

Abstract

Intracellular protein trafficking is the movement of membrane-bound organelles to and from requisite locations within the cell. Small GTPases are a critical component to the spatiotemporal accuracy of intracellular trafficking pathways as they determine the specificity and direction of organelle transport. There exists over 150 small GTPases categorized into 5 sub-families and are employed across all cell types. Despite their universal expression and relevance to cellular function, small GTPases remain incompletely understood across tissue types. In various instances, the trafficking pathway of a particular Rab in one cell type may belong to a completely disparate pathway in another cell type. Rab27 has been shown to traffic melanosomes in epithelial tissue, however, in endothelium, Rab27 has been linked to the trafficking of an endothelial specific vesicle known as the Weibel-Palade body (WPB). However, information is lacking as to whether Rab27 trafficking is applicable to sprouting angiogenesis. Rab35 is a well-studied Rab involved in a broad spectrum of cellular functions, including cytokinesis, cell migration, and cell polarity across tissue types. Surprisingly, Rab35 lacks investigation in endothelial cells. Like Rab35, Arf6 is also well-studied and implicated in several different cellular functions. It is most associated with cytoskeletal rearrangements, cell migration and endocytosis. Some Arf6 data exists in endothelial cells, but information is lacking on its role in sprouting angiogenesis and lumen formation. Rab8 is reported to traffic cargo exiting the trans-Golgi network bound for the apical membrane. The relevance of Rab8 to apical trafficking in endothelium remains unknown. Our data agrees that Rab27 is linked to the WPB pathway, and it is WPB cargo, Angiopoietin-2, that causes the hyper-sprouting phenotype observed in the absence of Rab27. In our hands, Rab35 does impact a range of endothelial cell processes. The extensive effects Rab35 induces is due to Rab35’s regulation of the actin cytoskeleton. We found that Arf6 also regulates the actin cytoskeleton but is critical to the management of transmembrane proteins important for sprouting angiogenesis and lumen formation. Finally, our results show that Rab8 is indeed important to apical trafficking in endothelium, independent of the WPB pathway. Together, these investigations provide insight to the divergent trafficking patterns of small GTPases in endothelium.

Publication Statement

Copyright is held by the author. User is responsible for all copyright compliance.

Rights Holder

Caitlin Francis

Provenance

Received from ProQuest

File Format

application/pdf

Language

en

File Size

245 pgs

Discipline

Biophysics

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