The main objective of this research project is to determine the signaling function of NEMOBP1 upon induction by the gram negative bacterial endotoxin, lypppolysacharide (LPS). LPS stimulates macrophages to express various inflammatory genes through their activation of toll-like receptors (TLRs). These TLRs elicit the signaling cascade via downstream kinases such as IkB kinase (IKK) and three families of MAP kinases (MARK): c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinases (ERK), and p38. IKK is a multi subunit complex composed of two catalytic subunits, IKKalpha and IKKbeta along with a regulatory subunit IKKgamma (also known as NEMO). Based on preliminary data obtained in Dr. Sun's laboratory, it is evident that NEMOBP1 physically interacts with the IKKy subunit and is also phosphorylated by IKK. In order to further understand the function of NEMOBP1, I plan to investigate the role of the NEMOBP1 in LPS-mediated induction of proinflammatory genes, such as TNF-alpha and IL-6. Secondly, I plan to determine the role of NEMOBP1 in mediating TLR signaling using bone marrow derived macrophages and peritoneal macrophages obtained from NEMOBP1 knockout and wild type mice. Lastly, I also plan to map the functional domains of NEMOBP1 by creating different N-terminal and C-terminal truncations along with some small internal deletions and point mutations of the probable functional regions. Based on my preliminary data, I believe that NEMOBP1 plays a critical role in cell signaling and also in the induction of proinflammatory genes.
