This project explores the function of a protein regulating release of neurotransmitters into synapses in the model organism Caenorhabditis elegans. A role for this protein in RNA processing is also explored. The gene unc-13 codes for protein products needed for priming synaptic vesicles before neurotransmitters are released into synapses. In Drosophila melanogaster, the levels of UNC-13 protein are regulated by degradation. Previous research identified an F-box protein and three proteins involved in RNA processing or expression that interact with C. elegans UNC-13. Aim 1) Mutations that suppress paralysis in unc-13 mutants will be identified. One such mutation was previously characterized in the PI's laboratory. This gene and other suppressors found in a screen will be identified. Aim 2) An F-box protein that interacts with UNC-13 and may be involved in degrading UNC-13 protein at synapses will be studied. Levels of UNC-13 will be examined after inhibition of degradation and following a decrease in expression of the F-box protein through double stranded RNA interference (RNAi). Proteins interacting with the F-box protein will be identified with the yeast two-hybrid system. Aim 3) Three proteins involved in RNA processing or expression that interact with UNC-13 will be examined. One of the proteins negatively regulates RNA interference, the second protein may be involved in silencing transgenes in the germline, and the third protein is involved in splicing mRNA. It will be determined whether the function of these proteins is altered in unc-13 mutants. In worms with decreased expression of these three proteins, expression and localization of UNC-13 will be examined.
BROADER IMPACT: Undergraduate students will be involved in all parts of the research project so that they will be better prepared for graduate school and may be more likely to pursue a career in research. Aim 1) All research projects described in this proposal will be carried out by undergraduate student researchers. Aim 2) Minority students in the Ursinus College Bridge Program will participate in experiments with C. elegans before they enroll in college. Two research positions will be available each summer for Bridge Program students completing their first year at Ursinus. A research collaboration and exchange will be maintained with a laboratory at Morgan State University, an Historically Black College/University (HBCU). Aim 3) A laboratory exercise in a case study based Molecular Neurobiology course integrating research in my laboratory will be enhanced. Aim 4) Training and C. elegans strains will be provided to a 'Science in Motion' outreach program to area public schools.
