Research Projects

The TBC (Tre-2/Bub2/Cdc16) domain-containing proteins are a family of proteins characterized by the presence of the conserved TBC (Tre-2/Bub2/Cdc16) domain. The TBC domain is approximately 200 amino acids long and serves as a catalytic domain with GTPase-activating protein (GAP) activity. TBC domain-containing proteins play a significant role in many biological processes including cancer, disease (Falace et al., 2010), cellular processes, and cell signaling (Kwon et al., 2017). A total of 40 TBC domains containing proteins have been identified in humans. Carabin also known as TBC1D10C is a GTPase activating protein (GAP) of Rab35, predominantly expressed in peripheral blood leukocytes and spleen. Carabin is a 446 amino acid long protein with a molecular weight of 47.7 kDa. It contains an N-terminal Rab-GTPase binding domain and a C-terminal calcineurin binding site and has been shown to inhibit Ras and calcineurin pathways during T cell activation. Additionally, Carabin has been found to inhibit B cells activation by targeting Ras. Increased expression of Carabin in macrophages has been shown to result in higher phagocytic activity against Burkholderia cenocepacia. Conversely, low levels of Carabin expression have been observed in patients with systemic lupus erythematosus (SLE), suggesting its involvement in bacterial infections and autoimmunity. Despite its known functions in T cells, B cells, and macrophages, its role in gut (intestinal tissue ) where its expression is higher after bone marrow and lymphoid tissue (HPA data analysis, Fig-1), remains unexplored. The intestinal epithelium serves as the barrier against pathogenic bacterial infections. In this study, we aim to investigate the functional role of Carabin in regulating intestinal epithelial cell (IEC) function during pathogenic bacterial infections. We will utilize pathogenic bacteria such as Salmonella typhimurium or E. coli (EHEC) to infect IEC lines (SW480, HT-29 or Caco-2) wild type and Carabin knockdown/knockout to elucidate the impact of Carabin on IECs functions. Our study will focus on determining whether Carabin modulates inflammation and regulates the autophagy process to control inflammation as well as its potential role in restricting intracellular bacterial growth. To comprehensively understand the global response and underlying mechanisms of Carabin in IECs, we will conduct RNA sequencing. Additionally, we will perform immunoprecipitation assays and mass spectrometry to identify Carabin’s interacting partners.