Title page for E-project-121505-113309

Project Type MQP

Submission date 2005-12-15

Author
Jamie B Towle, BIO

URN E-project-121505-113309

Sponsor UMass Medical School

Title Exploring the Interactions of Yeast Exocyst Complex Proteins Using Immunoprecipitation Experiments

Advisor
Adams, David S., BB

Availability unrestricted

Abstract
Vesicle trafficking is vital for the growth and life of a cell. The exocyst is an eight protein complex involved in vesicular trafficking. A model of the interactions among the exocyst proteins was proposed through yeast 2-hybrid and in vitro translation experiments, but has not been demonstrated by more stringent methods. To support these findings in vivo and to further map the binding domains, a series of immunoprecipitation experiments was performed. Green fluorescent protein (GFP) genomically tagged proteins Sec5p and Sec8p were pulled down from whole yeast lysates, and the samples were probed with anti-Sec6 antibody. If Sec6p binds Sec5-GFP or Sec8-GFP a band will appear in the western blot in the bound lane. Likewise, a similar experiment was carried out using Sec8-myc to determine its interaction with Sec6p. Results from both experiments show an in vivo interaction between Sec6p and Sec5-GFP and Sec8-GFP.

Files
MQP.pdf

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Project Type	MQP
Submission date	2005-12-15
Author	Jamie B Towle, BIO
URN	E-project-121505-113309
Sponsor	UMass Medical School
Title	Exploring the Interactions of Yeast Exocyst Complex Proteins Using Immunoprecipitation Experiments
Advisor	Adams, David S., BB
Availability	unrestricted
Abstract Vesicle trafficking is vital for the growth and life of a cell. The exocyst is an eight protein complex involved in vesicular trafficking. A model of the interactions among the exocyst proteins was proposed through yeast 2-hybrid and in vitro translation experiments, but has not been demonstrated by more stringent methods. To support these findings in vivo and to further map the binding domains, a series of immunoprecipitation experiments was performed. Green fluorescent protein (GFP) genomically tagged proteins Sec5p and Sec8p were pulled down from whole yeast lysates, and the samples were probed with anti-Sec6 antibody. If Sec6p binds Sec5-GFP or Sec8-GFP a band will appear in the western blot in the bound lane. Likewise, a similar experiment was carried out using Sec8-myc to determine its interaction with Sec6p. Results from both experiments show an in vivo interaction between Sec6p and Sec5-GFP and Sec8-GFP.
Files	MQP.pdf