The overall aim of the proposed research is to understand the role of paternally contributed components for the earliest stages of animal development. Paternal effect mutants provide powerful tools to identify molecules of paternal origin that are essential for embryogenesis. The function of a set of genes whose products act within a narrow window of time, from the initial response of the sperm to the egg cytoplasm to the completion of the first embryonic mitosis, will be studied at the cellular, genetic, and molecular levels.
SPECIFIC AIMS
1) To understand the function of the ms(3)sneaky (snky) gene in sperm activation after entry into the egg, the Wakimoto lab will:
a) molecularly clone the gene to characterize its expression profile and identify its RNA and protein products b) determine the subcellular distribution of the SNKY protein by immunolocalization c) track the behavior of membrane markers in snky mutant sperm to test the hypothesis that the paternal effect defect results from failure of plasma membrane breakdown
2) To understand the role of the ms(3)K81 gene in restructuring or replicating the sperm nucleus after insemination, the lab will:
a) examine the distribution of the K81 transcript and protein during spermatogenesis and embryogenesis using immunolocalization b) examine the functional relatedness of K81 to a newly discovered, closely related K81-like Drosophila protein
3) To examine the functional relationships between genes that are similar to snky and-K81 in causing early developmental defects; the lab will:
a) genetically characterize and initiate molecular mapping of a newly discovered gene called pop-sickle, which when mutant is phenotypically indistinguishable from the snky mutation b) genetically characterize and initiate molecular mapping of a newly discovered gene called deadbeat, which when mutant is phenotypically indistinguishable from the K81 mutation c) examine the genetic interactions between these genes, snky and K81 to elucidate their relative times of action
