Abstract
Adenovirus is a nonenveloped virus that consists of an icosahedral capsid that contains double-stranded DNA. Viruses of this type can cause the common cold, conjunctivitis, and diarrhea. An important gene of adenoviruses is an early viral gene called E4 ORF3, which encodes the protein, E4 11k. This gene is responsible for changing the cellular environment, which aids in viral replication and prevents the host cell’s synthesis of proteins. E4 11k binds to Ddx6, a cellular protein located in P-bodies, and disrupts its localization. P-bodies are granules in the cytoplasm that contain mRNAs that have been repressed and proteins that are related to the degradation of mRNA. Overall, the purpose of this experiment is to determine the binding site of E4 11k and its interaction with Ddx6. This research will be conducted by transfecting cells with Ddx6 and E4 11k mutants and determining their binding ability. To begin, the transfection efficiency must be optimized. This was performed by observing protein concentrations of the transfected genes by immunoblotting. In addition, the single-cell concentrations were examined through immunofluorescence. This innovative research will eventually aid in learning about the effect of the interaction between E4 11k and Ddx6 on the viral life cycle.
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The Binding Site Between the Adenoviral Protein E4 11k and the Cellular Protein Ddx6
Adenovirus is a nonenveloped virus that consists of an icosahedral capsid that contains double-stranded DNA. Viruses of this type can cause the common cold, conjunctivitis, and diarrhea. An important gene of adenoviruses is an early viral gene called E4 ORF3, which encodes the protein, E4 11k. This gene is responsible for changing the cellular environment, which aids in viral replication and prevents the host cell’s synthesis of proteins. E4 11k binds to Ddx6, a cellular protein located in P-bodies, and disrupts its localization. P-bodies are granules in the cytoplasm that contain mRNAs that have been repressed and proteins that are related to the degradation of mRNA. Overall, the purpose of this experiment is to determine the binding site of E4 11k and its interaction with Ddx6. This research will be conducted by transfecting cells with Ddx6 and E4 11k mutants and determining their binding ability. To begin, the transfection efficiency must be optimized. This was performed by observing protein concentrations of the transfected genes by immunoblotting. In addition, the single-cell concentrations were examined through immunofluorescence. This innovative research will eventually aid in learning about the effect of the interaction between E4 11k and Ddx6 on the viral life cycle.