Author

Uma Saimani

Date of Graduation

Fall 2016

Degree

Master of Science in Biology

Department

Biology

Committee Chair

Kyoungtae Kim

Keywords

Vps1, Vps51, retrograde pathway, endosome, TGN, GARP, yeast

Subject Categories

Biology

Abstract

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 replacement of these residues led to severe defects in endosome-to-TGN transport of Snc1, providing evidence of the physiological relevance of the interaction of Vps51 with Vps1 via these residues for the traffic. Furthermore, my functional analysis revealed that Vps1 acts upstream of Vps51, and the absence of Vps1 resulted in reduced localization levels of Vps51 and its binding partner Tlg1 to the late Golgi. Taken together, I propose that Vps1 functions with the GARP tethering machinery for efficient tethering/fusion at the TGN.

Copyright

© Uma Saimani

Open Access

Included in

Biology Commons

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