Date of Graduation

Spring 2014

Degree

Master of Science in Biology

Department

Biology

Committee Chair

Kyoungtae Kim

Abstract

Vps1 (vacuole protein sorting) is a GTPase protein and a yeast ortholog to the mammalian dynamin-like protein (Dlp1). Vps1 acts cooperatively with many proteins at diverse cellular locations for endocytosis, protein sorting, and membrane fusion and fission. Classical dynamin family proteins work in concert with clathrin to facilitate not only receptor-mediated endocytosis, but also traffic from the Golgi to endosomes. In this study, the interdependency between Vps1 and clathrin was investigated. Subcellular localization of clathrin was at the Golgi, early and late endosomes, implicating clathrin in multiple trafficking pathways within the cell. Vps1 and clathrin colocalize, and loss of Vps1 led to a shift in the cellular localization of clathrin to the late endosome and vacuole. However, loss of clathrin had no effect on the localization of Vps1. In addition, Golgi PtdIns(4)P is increased upon loss of Vps1, supporting a role for Vps1 as a pinchase of the Golgi membrane. The physical interaction between Vps1 and clathrin was examined using a Yeast Two-Hybrid system, and found that Vps1 and clathrin indeed interact in vivo. Together, these findings demonstrate a functional relationship between Vps1 and clathrin, which is consistent with previous models whereby Vps1 mediates scission of clathrin coated vesicles at the Trans-Golgi network.

Keywords

Vps1, clathrin, Golgi, dynamin, dynamin-like protein

Subject Categories

Biology

Copyright

© Michelle Amber Williams

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