Abnormalities of protein function within the central nervous system contribute to diseases of cognitive function, motor function, and development in children and adults. One method to determine the function of a protein is to identify proteins with which it interacts. Yeast two hybrid assays have been successful for identifying protein-protein interactions of brain specific proteins important in synapse formation, neurotransmitter release, and receptor signaling. Studying networks of protein-protein interactions of the brain will lead to a better understanding of neurological diseases with complex genotypes and multiple phenotypes. In addition, these proteomic and network analyses should reveal pathways that are amenable to diagnostics and to drug targeting for therapeutic purposes.

The traditional yeast two hybrid system relies on two chimeric proteins often referred to as bait and prey. One fusion protein contains a gal-4-DNA binding domain and gene of interest. The other protein is a fusion of a transcriptional activator and a protein domain derived from a cDNA library. If two proteins interact, the DNA binding fusion protein is brought into contact with the activation domain fusion protein and the output signal is the induction of a reporter gene. Colonies of yeast with reporter gene activity are then analyzed by PCR to determine the interacting protein sequence.

  • Yeast two hybrid Screens: The core is presently using Invitrogen Gateway technology to screen proteins of interest to NINDS researchers at UAB.