Do Drosophila nuage-colocalizing components, Vasa and Maelstrom, interact?
Drosophila oogenesis is the process where a stem cell becomes a mature oocyte; a single large cell in which both body axes are established in the form of asymmetrically distributed morphogens. Oogenesis sets the stage for the next phase in development called embryogenesis, which produces the larva and ultimately the adult. For most of oogenesis, the oocyte develops within the context of a continuous cytoplasm with 15 other germline cells, called nurse cells, whose function is to provide virtually all gene products required over the course of oogenesis. A novel Drosophila protein, Maelstrom, and an RNA helicase, Vasa, colocalize for most of oogenesis to a cryptic subcellular structure called the nuage. The nuage is a hallmark feature of cells of the germ line lineage across the animal kingdom, from planaria to vertebrates; and is therefore implicated in the germline cell function. Recent experiments have revealed phenotypic similarity between mutants of these respective genes. Based on similarity of phenotype and localization pattern, an imperative question then is whether or not Vasa and Maelstrom interact physically. To this end, the first part of my project will be to perform GST protein pull down assays to detect any binding between the two proteins, in vitro. The second part of my project will be aimed toward characterizing the nature of the nuage. Therefore, additional experiments using biochemical techniques such as sucrose gradients will be used to help identify additional components that are involved in this interaction.
