Using the Auxin-Induced Degradation System to Dissect Factors Within the Exocyst

Heard, Rachael Denielle

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Using the Auxin-Induced Degradation System to Dissect Factors Within the Exocyst

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I am interested in understanding the roles of different subunits in controlling the polarized localization of exocyst. Specifically, I asked what happens to the structure, stability and localization of exocyst when other subunits are degraded. An auxin induced degradation system (AID) will be used with the subunits Sec3, Sec6, and Exo70 to determine if the structure of the complex is affected by this degradation. The AID system allows for rapid depletion of targeted proteins in response to the hormone auxin. I took advantage of the GFP tag on Sec8 in order to determine what subunits are still attached to Sec8 when degradation occurs, by pulling down the complex onto GFP bound nanobody beads. Next, I determined where Sec8-GFP is localized in the cell after Sec6, Sec3, and Exo70 degradation.

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