The molecular motor cytoplasmic dynein is essential for many aspects of cellular functioning, including organelle transport, mitosis and cell division. Molecular motors are important in all cells, but they are of particular importance in irons. Mutations in motor proteins cause severe neuronal impairment and many neurodegenerative diseases exhibit failures in organelle transport. The range of functions of molecular motors makes their regulation a critical question. A significant facet of this regulation is the targeting of specific cargoes to the motor complex. Recently, several light chains ~e been identified which may be involved in the motor/cargo interactions of cytoplasmic dynein. One hypothesis is that rein complexes may contain different light chains, or different combinations of light chains, and these differences may be solved in targeting the appropriate cargo to the motor complex. This project seeks to understand the role of dynein light bins in motor/cargo interactions. Affinity columns will be used to identify neuronal proteins that bind to each light chain. Imunocytochemistry will be used to determine if the identified proteins co- localize to the same cellular compartments and immunoprecipitation will be used to determine if the proteins interact with cytoplasmic dynein in vivo. Finally, green flourescent and red fluorescent tagged constructs of the identified proteins and dynein light chains will be constructed to examine their interactions and dynamics in living cells.
