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Full-Text Articles in Life Sciences

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 …


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 …