The long term goals of this project are to identify genes which directly affect the longevity of Drosophila, and ultimately, mammals. The candidates for this function in Drosophila are the heat shock genes, and a number of other genes which have been found to be differentially expressed at the transcriptional level during aging. Any correlation of their expression with longevity will be determined by examining RNA levels in strains of flies which have been genetically selected for postponed aging ("O" strains), and their controls ("B" strains). A direct role in regulating longevity will then be assayed by experimentally increasing and decreasing the expression of the genes in transgenic flies. These experiments will involve the development of new vectors for overexpression of candidate LAGs, and the creation of stocks exhibiting postponed aging which are compatible with transgenic technologies. Finally, any mammalian homologs of Drosophila LAGs will be identified. The specific aims are: 1. Analyze heat shock gene RNA expression in existing O & B strains. 2. Analyze expression of additional cloned candidate LAGs in existing O & B strains. 3. Create ry506 and w1118 marked O and B strains: O[ry] & B[ry], and Ow & Bw. 4. Create and test new inducible-overexpression transformation vectors, and overexpress candidate LAGs in Drosophila strains, to analyze effects on longevity. 5. Underexpress candidate LAGs in Drosophila strains, and analyze longevity. 6. Introduce selected transformation constructs, and enhancer-traps with aging-specific expression patterns, into new marked O[ry] and B[ry] strains. 7. Identify and clone rodent and human homologs of Drosophila LAGs.