Initial Studies in the Interaction Between the Cellular Ddx6 and Adenovirus E4 orf3 Proteins
Abstract
Adenoviruses cause a number of diseases including many that infect the respiratory tract, leading to the common cold, along with those that infect the conjunctiva and the gastrointestinal tract. E4 orf 3, an adenovirus protein, has been shown to directly bind to the Ddx6 protein of the cellular P-body. P-bodies play a role in mRNA regulation and the interaction of Ddx6 and E4 orf3 may be involved in increasing viral late gene expression. The aim of this experiment is to narrow down the binding site between the two in order to create mutants that will help determine the function of this interaction. Using E4 orf 3 and N- and C-terminal deletion mutants of the P-body protein, Ddx6, co-immunoprecipitation techniques will be employed to determine which end of Ddx6 contains the region of binding for E4 orf3. Before these results can be achieved, we need sufficient expression of these proteins in human cells so the most efficient protocol for transfection is being assessed. Two transfection techniques, CaPO4 and Lipofectamine 3000, are being tested to determine which results in a more effective transfection of the expression vectors into the host cells. Because adenovirus serves as an effective model for other families of viruses, the interactions between E4 orf 3 and Ddx6 will be able to show us more about how P-bodies interact with other viruses that may cause more detrimental diseases.
Session Name:
Biology II
Start Date
10-4-2015 10:15 AM
End Date
10-4-2015 11:15 AM
Location
HSB 207
Initial Studies in the Interaction Between the Cellular Ddx6 and Adenovirus E4 orf3 Proteins
HSB 207
Adenoviruses cause a number of diseases including many that infect the respiratory tract, leading to the common cold, along with those that infect the conjunctiva and the gastrointestinal tract. E4 orf 3, an adenovirus protein, has been shown to directly bind to the Ddx6 protein of the cellular P-body. P-bodies play a role in mRNA regulation and the interaction of Ddx6 and E4 orf3 may be involved in increasing viral late gene expression. The aim of this experiment is to narrow down the binding site between the two in order to create mutants that will help determine the function of this interaction. Using E4 orf 3 and N- and C-terminal deletion mutants of the P-body protein, Ddx6, co-immunoprecipitation techniques will be employed to determine which end of Ddx6 contains the region of binding for E4 orf3. Before these results can be achieved, we need sufficient expression of these proteins in human cells so the most efficient protocol for transfection is being assessed. Two transfection techniques, CaPO4 and Lipofectamine 3000, are being tested to determine which results in a more effective transfection of the expression vectors into the host cells. Because adenovirus serves as an effective model for other families of viruses, the interactions between E4 orf 3 and Ddx6 will be able to show us more about how P-bodies interact with other viruses that may cause more detrimental diseases.