Neisseria gonorrhoeae (the gonococcus) causes the sexually transmitted disease gonorrhea. About 400,000 cases are reported in the U.S. annually. Opacity (Opa) proteins on the surface of the gonococci promote adherence and entry into human epithelial cells and neutrophils. The focus of this proposal is to use molecular genetic approaches, specifically the yeast two-hybrid system, to dissect interactions between Opa and 2 Opa interacting proteins (OIPs). Using deletions of Opa, domains of the protein interacting with OIP1 and OIP3 will be determined using the yeast two-hybrid system. OIP1 contains three LIM domains thought to be important in protein-protein interactions. Constructs containing individual LIM domains will be generated and tested to see which LIM domain is responsible for interaction with Opa. Bacterial adherence and invasion studies will be performed to begin to address the role of OIP1 in intracellular survival of gonococci. In addition, immunofluorescence assays will be done to verify interactions between OIP1 and Opa in vivo.
