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Full-Text Articles in Life Sciences
Multifaceted Mechanism Of Vps1 Mediated Endosome-To-Golgi Fusion In Vitro, Ehsan Suez
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
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 …
Viral Fusion Protein Tm-Tm Interactions: Modulators Of Protein Function And Potential Antiviral Targets, Stacy Webb
Viral Fusion Protein Tm-Tm Interactions: Modulators Of Protein Function And Potential Antiviral Targets, Stacy Webb
Theses and Dissertations--Molecular and Cellular Biochemistry
Enveloped viruses, such as HIV, influenza, and Ebola, utilize surface glycoproteins to bind and fuse with a target cell membrane. This fusion event is necessary for release of viral genomic material so the virus can ultimately reproduce and spread. The recently emerged Hendra virus (HeV) is a negative-sense, single-stranded RNA paramyxovirus that presents a considerable threat to human health as there are currently no human vaccines or antivirals available. The HeV utilizes two surface glycoproteins, the fusion protein (F) and the attachment protein (G), to drive membrane fusion. Through this process, the F protein undergoes an irreversible conformational change, transitioning …
Membrane Fusion Proteins Are Required For Oskar Mrna Localization In The Drosophila Egg Chamber, Douglas Ruden, Vincent Sollars, Xiaoyan Wang, Daisuke Mori, Marina Alterman, Xiangyi Lu
Membrane Fusion Proteins Are Required For Oskar Mrna Localization In The Drosophila Egg Chamber, Douglas Ruden, Vincent Sollars, Xiaoyan Wang, Daisuke Mori, Marina Alterman, Xiangyi Lu
Vincent E Sollars
We used a genetic screen in Drosophila to identify mutations which disrupt the localization of oskar mRNA during oogenesis. Based on the hypothesis that some cytoskeletal components which are required during the mitotic divisions will also be required for oskar mRNA localization during oogenesis, we designed the following genetic screen. We screened for P-element insertions in genes which slow down the blastoderm mitotic divisions. A secondary genetic screen was to generate female germ-line clones of these potential cell division cycle genes and to identify those which cause the mislocalization of oskar mRNA. We identified mutations in ter94 which disrupt the …
A Lipid-Anchored Snare Supports Membrane Fusion, Hao Xu, Michael Zick, William T. Wickner, Youngsoo Jun
A Lipid-Anchored Snare Supports Membrane Fusion, Hao Xu, Michael Zick, William T. Wickner, Youngsoo Jun
Dartmouth Scholarship
Intracellular membrane fusion requires R-SNAREs and Q-SNAREs to assemble into a four-helical parallel coiled-coil, with their hydrophobic anchors spanning the two apposed membranes. Based on the fusion properties of chemically defined SNARE- proteoliposomes, it has been proposed that the assembly of this helical bundle transduces force through the entire bilayer via the transmembrane SNARE anchor domains to drive fusion. However, an R-SNARE, Nyv1p, with a genetically engineered lipid anchor that spans half of the bilayer suffices for the fusion of isolated vacuoles, although this organelle has other R-SNAREs. To demonstrate unequivocally the fusion activity of lipid-anchored Nyv1p, we reconstituted proteoliposomes …
Resolution Of Organelle Docking And Fusion Kinetics In A Cell-Free Assay, Alexey J. Merz, William T. Wickner
Resolution Of Organelle Docking And Fusion Kinetics In A Cell-Free Assay, Alexey J. Merz, William T. Wickner
Dartmouth Scholarship
In vitro assays of compartment mixing have been key tools in the biochemical dissection of organelle docking and fusion. Many such assays measure compartment mixing through the enzymatic modification of reporter proteins. Homotypic fusion of yeast vacuoles is measured with a coupled assay of proteolytic maturation of pro-alkaline phosphatase (pro-ALP). A kinetic lag is observed between the end of docking, marked by the acquisition of resistance to anti-SNARE reagents, and ALP maturation. We therefore asked whether the time taken for pro-ALP maturation adds a kinetic lag to the measured fusion signal. Prb1p promotes ALP maturation; overproduction of Prb1p accelerates ALP …