Project description:Newcastle Disease Virus (NDV) is one of the most threatening viruses to the poultry industry, and it also exhibits oncolytic properties. In our research, we identified a non-oncolytic strain, genotype VII NDV strain I4, where the lack of oncolytic ability is attributed to its NP protein. To explore the mechanism of NP-mediated oncolysis by NDV, we focused on the differential interacting proteins of the NP proteins from two strains: the highly oncolytic Herts/33 strain and the poorly oncolytic I4 strain. The mass spectrometry results show the interacting protein profiles of the NP proteins from both virus strains. After infecting HeLa cells with each strain, we conducted immunoprecipitation using antibodies against the NP protein. Due to the lower expression of I4-associated proteins during replication, we further performed immunoprecipitation experiments using separately overexpressed NP proteins from I4 and Herts/33.
Project description:Newcastle disease virus (NDV) is one of the most significant threats to the poultry industry, and it also exhibits oncolytic properties. In our research, we have identified a non-oncolytic strain, the genotype VII NDV strain I4. We have found that the lack of oncolytic characteristics in this strain is attributed to amino acids 366-489 in the NP protein. To delve into the mechanism of NP-mediated NDV oncolysis, we initiated further investigations using the highly oncolytic Herts/33 strain's NP and a recombinant NP protein with amino acids 366-489 from the I4 strain substituted in (H-NP366-489I). These proteins represent the interacting partners in the proteomics analysis of NP proteins from both viral strains. After infecting HeLa cells with each of these strains, immunoprecipitation was performed using antibodies targeting the NP protein. Since the H-NP366-489I virus produces fewer proteins during replication, we additionally overexpressed the H-NP366-489I and Herts/33 NP proteins separately for immunoprecipitation experiments.
Project description:Purpose of study was to investigate whole genome expression changes of a strain with deletion of the two-component system TctD-TctE and determine genes dysregulate relative to the parental wildtype to gain insight into possible regulatory targets of TctD-TctE. TctD-TctE is a two-component system in Pseudomonas aeruginosa that responds to and regulates uptake of tricarboxylic acids such as citric acid. It accomplishes this through derepression of the porin encoding the gene opdH, thereby regulating influx of citrate metabolites from the surrounding environment. Deletion of the tctED operon (ΔtctED) resulted in a reduced growth phenotype when citric acid is present in media. In the ΔtctED strain the presence of citric acid was found to have an inhibitory effect on growth. When the alternative carbon source arginine was present, wildtype levels of growth could not be restored. Static cultures of ΔtctED had low cell density in the presence of citric acid but maintained the same levels of biofilm formation compared to conditions when no citric acid was present. This suggests a dysregulation of biofilm formation in the presence of citric acid. In the ΔtctED strain there was also greater accumulation of tobramycin within the biofilm compared to the PA14 wildtype strain. Additionally, analysis of whole-genome expression found that multiple metabolic genes were dysregulated in ΔtctED. Here it is concluded that TctD-TctE is involved in biofilm tolerance to tobramycin in the presence of citrate metabolites.