Presentation Title

Investigating the Binding of Spire/Myosin-V Binding in Drosophila melanogaster

Faculty Mentor

Dr. Margot E. Quinlan

Start Date

23-11-2019 8:45 AM

End Date

23-11-2019 9:30 AM

Location

64

Session

poster 2

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Spire works with Cappuccino to build actin filaments in the cell. Both proteins play an important role in polarity establishment in Drosophila oocytes by constructing the actin meshwork. Myosin V is a motor protein that travels along actin filaments like those that Spire and Cappuccino construct. Previous research suggests that Myosin V and Spire interact. We hypothesize that Myosin V is activated by Spire, but that the binding of Myosin V and Spire has no effect on Spire’s activity. In order to test this, we purified the two proteins from E. coli and tested their binding in a pulldown assay. The purpose of this study is to measure binding of Myosin V and Spire and learn how it affects each of their functions. In order to test for binding of Spire and Myosin V, we performed a pulldown assay with wild type and mutated Spire and Myosin V. Results suggest when either Spire or Myosin V are mutated, binding is greatly reduced compared to binding of wild type Spire and wild type Myosin V. This shows that mutating either Spire or Myosin V affects binding of the two proteins. Analysis of Spire’s actin assembly in vitro coupled with in vivo data using the Drosophila oocyte as a system will reveal the importance of the Spire-Myosin V interaction. We predict that the disruption of binding between Spire and Myosin V will negatively affect viability and meshwork formation in Drosophila oocytes.

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Nov 23rd, 8:45 AM Nov 23rd, 9:30 AM

Investigating the Binding of Spire/Myosin-V Binding in Drosophila melanogaster

64

Spire works with Cappuccino to build actin filaments in the cell. Both proteins play an important role in polarity establishment in Drosophila oocytes by constructing the actin meshwork. Myosin V is a motor protein that travels along actin filaments like those that Spire and Cappuccino construct. Previous research suggests that Myosin V and Spire interact. We hypothesize that Myosin V is activated by Spire, but that the binding of Myosin V and Spire has no effect on Spire’s activity. In order to test this, we purified the two proteins from E. coli and tested their binding in a pulldown assay. The purpose of this study is to measure binding of Myosin V and Spire and learn how it affects each of their functions. In order to test for binding of Spire and Myosin V, we performed a pulldown assay with wild type and mutated Spire and Myosin V. Results suggest when either Spire or Myosin V are mutated, binding is greatly reduced compared to binding of wild type Spire and wild type Myosin V. This shows that mutating either Spire or Myosin V affects binding of the two proteins. Analysis of Spire’s actin assembly in vitro coupled with in vivo data using the Drosophila oocyte as a system will reveal the importance of the Spire-Myosin V interaction. We predict that the disruption of binding between Spire and Myosin V will negatively affect viability and meshwork formation in Drosophila oocytes.