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Full-Text Articles in Life Sciences

Multifaceted Mechanism Of Vps1 Mediated Endosome-To-Golgi Fusion In Vitro, Ehsan Suez Aug 2020

Multifaceted Mechanism Of Vps1 Mediated Endosome-To-Golgi Fusion In Vitro, Ehsan Suez

MSU Graduate Theses

To maintain cell homeostasis, protein recycling through intracellular membrane fusion is an important cellular process. Both Vps1 and Ypt6 have been implicated in protein recycling from the endosome to the trans-Golgi Network (TGN). SNARE proteins are thought to be the key regulator in this membrane fusion mediated protein recycling mechanism. I studied membrane fusion events by incorporating purified proteins into liposomes. A series of data suggest that high concentration of SNARE proteins inhibits fusion unlike the opposite popular notion. Also, the data suggests that Vps1 acts on membrane fusion dynamics in a manner that lower concentrations of Vps1 enhance …


The Dynamin-Like Protein Vps1 Stimulates Endosome-To-Golgi Fusion In Vitro, Jared Christopher Smothers Aug 2019

The Dynamin-Like Protein Vps1 Stimulates Endosome-To-Golgi Fusion In Vitro, Jared Christopher Smothers

MSU Graduate Theses

Intracellular membrane fusion events can be reconstituted by exploiting isolated organelles from cellular hosts or artificial membranes made of purified phospholipid components. Artificial construction of membranes provides two significant advantages. First, cellular isolation of the endosome-derived vesicles and TGN (trans-Golgi Network) compartments needed for the fusion assay would be extremely challenging. Second, reconstituting the membranes provides the added benefit of controlling size and lipid compositions to functionally mimic the individual membrane architectures and introduce only the purified proteins that are under investigation. For these reasons, I have developed the first simultaneous lipid and content mixing fusion assays that measures the …


Yeast Dynamin And Ypt6 Converge On The Garp For Endosome-To-Golgi Trafficking, Pelin Makaraci Dec 2017

Yeast Dynamin And Ypt6 Converge On The Garp For Endosome-To-Golgi Trafficking, Pelin Makaraci

MSU Graduate Theses

Protein recycling is an important cellular process required for cell homeostasis. Results from prior studies demonstrated that Vps1, a dynamin homologue in yeast, is implicated in protein recycling from the endosome to the trans-Golgi Network (TGN). However, the function of Vps1 in relation to Ypt6, a master GTPase in the recycling pathway, remains unknown. The present study reveals that Vps1 physically interacts with Ypt6 if at least one of them is full-length. It was found that overexpression of full-length Vps1, but not GTP hydrolysis-defective Vps1 mutants, is sufficient to rescue abnormal phenotypes in membrane trafficking pathways provoked by loss …


Yeast Dynamin Associates With The Garp Tethering Complex For Endosome-To-Golgi Traffic, Uma Saimani Dec 2016

Yeast Dynamin Associates With The Garp Tethering Complex For Endosome-To-Golgi Traffic, Uma Saimani

MSU Graduate Theses

Yeast dynamin, Vacuolar Protein Sorting 1 (Vps1), has been implicated in recycling proteins and lipids from the endosome to the trans-Golgi network (TGN). Previous research showed a genetic interaction of Vps1 with all components of the GARP tethering complex, which anchors vesicles at the late Golgi membrane. I used the yeast two-hybrid system and pinpointed a 33 amino acid segment of Vps51, a GARP subunit, that interacts with Vps1. Based on sequence homology between Vps51 and its mammalian homolog ANG2 in the 33 amino acid stretch, I identified two key residues of Vps51, E127, and Y129, that bind Vps1. The …


Yeast Dynamin Functions With Escrt-Ii At The Late Endosome And Potential Roles With Novel Binding Partners, Bryan Thien Banh Jan 2016

Yeast Dynamin Functions With Escrt-Ii At The Late Endosome And Potential Roles With Novel Binding Partners, Bryan Thien Banh

MSU Graduate Theses

Yeast dynamin, Vps1, is a dynamic protein implicated in multiple trafficking pathways and at several cellular locations including the Golgi, endosome, and vacuole. Previous studies have found that Vps1 genetically and physically interacts with several ESCRT-II (Vps22, Vps36) and ESCRT-III (Vps2, Vps24) components. In light of these findings, little is known about how Vps1 functions with the ESCRT system. In my study, I have mapped the interaction domains between Vps1 and Vps22/Vps36 as well as explored the potential roles of these proteins during the targeting of vacuolar hydrolase, Cps1. My results support the idea of Vps1 working at the endosomal …


Dynamin Association With Clathrin And Its Physiological Roles At The Golgi And Targeting Mechanism To The Golgi, Shiva Kumar Goud Gadila Jan 2015

Dynamin Association With Clathrin And Its Physiological Roles At The Golgi And Targeting Mechanism To The Golgi, Shiva Kumar Goud Gadila

MSU Graduate Theses

Vps1 (Vacuole protein sorting 1), a dynamin-like protein in yeast, is implicated in diverse membrane trafficking pathways and localized to many organelles, including the Golgi. It has been proposed that Vps1 is functionally linked to clathrin heavy chain 1 (Chc1), but the question of how, where, and when they function together remains unknown. Lines of evidence suggest that Chc1 and Vps1 are located at the late Golgi. Therefore, I hypothesized that Vps1 binds to Chc1 and functions together at the Golgi for efficient Golgi-to-endosome membrane trafficking. Using the yeast-two hybrid assay, I present evidence that Vps1 binds to the C-terminal …


Functional Connection Between Yeast Dynamin And Retromer At The Endosome, Christopher Robert Trousdale Jan 2015

Functional Connection Between Yeast Dynamin And Retromer At The Endosome, Christopher Robert Trousdale

MSU Graduate Theses

Intracellular trafficking from the late endosome to Golgi in cells is termed retrograde transport, essential for recycling of important macromolecules including cell membrane receptors. Retrograde transport is regulated by a family of proteins known as the "Retromer” composed of 5 VPS proteins (Vps5, Vps17, Vps26, Vps29, and Vps35). Retromer acts as the coat proteins for vesicles emerging from late endosomes. Loss of Retromer function has been previously implicated in both Parkinson's and Alzheimer's disease. Vps1, a yeast dynamin-like protein, plays a role in intracellular trafficking. Vps1 has been shown to localize at the endocytic sites to promote pinching off of …