Long term work in Dr. T. H. Dunnebacke's laboratory, provocative but generally ignored, has shown that an agent from amoebae of Naegleria gruberi induces generations-delayed death of vertebrate cells in culture, and that these dying cells in turn produce an agent that causes death of other vertebrate cells. This remarkable phenomenon has been confirmed, and it has been shown that the cell death has features characteristic of apoptosis. The first goal is to complete and carefully document these new observations. Dunnebacke has purified the agent from amoebae, and has identified it as a protein, which has been partially sequenced. The plan, in collaboration with Dunnebacke, is to use her information to clone the Naegleria gene that encodes the protein, to sequence and characterize this gene and its product, and to determine whether the isolated protein is responsible for the biological activity of the agent. These studies should make this phenomenon a paradigm for future investigations of: 1, the mechanism by which a protein induces generations-delayed apoptosis; 2, how a protein from an amoeba can lead to the production, by dying vertebrate cells, of a product with similar biological activity, as well as the nature of the vertebrate product; and 3, possible therapeutic applications of the protein.
