Project description:Infection caused by bacteria from environmental reservoirs such as E. coli and S. uberis have not decreased in prevalence. Lack of success in controlling bovine mastitis due to S. uberis is associated with the route of infection which is not well understood and there is inadequate information on pathogenesis of S. uberis. Therefore, this study was to investigate the virulence factors of S. uberis using comparative genome analyses using isolates from cows with clinical mastitis and isolates from cows with a low cell count in their milk using a Subtracted Diversity Array (SDA). This study also reports the construction and validation of a microarray capable of fingerprinting the virulent and non-virulent isolates using the SDA technique.
Project description:Herein, we explore the druggability of BAG-1S (the smallest isoform) with a specific emphasis on its interaction with C-RAF. We firstly characterize the higher order structure of BAG-1S structure to determine druggable sites by HDX-MS. We then map the BAG-1S:c-Raf interface – identifying a 20 amino acid region likely to interact with c-Raf. Following which, we use mutational analysis of BAG-1S to show specific amino acids that disrupt the BAG-1S:c-Raf interaction, downstream signaling and cell growth. Finally, we develop a BAG-1S interacting peptide (from c-Raf) that disrupts BAG-1 protein:protein interactions (including C/B-RAF, CHIP and HSP70) and modified with a cell-penetrating motif which inhibits the growth of cancer cells with associated signaling inhibition and induction of apoptosis.