The long term objective of the project is to study the polypeptide structure of the nuclear pore complexes in Drosophila melanogaster. Work will focus on a myosin-like ATPase recently localized to Drosophila nuclear pore complexes and thought to participate in nucleocytoplasmic transport. The specific goal of this proposal is to biochemically identify the nuclear myosin-like protein. Work will concentrate on the purification of this protein from Drosophila Kc tissue culture cells and/or early embryos using affinity chromatography strategies. Once the nuclear myosin-like ATPase protein has been affinity purified, amino acid sequence information will be used to generate monospecific antibodies and oligonucleotides to clone and sequence the gene encoding the Drosophila nuclear myosin-like ATPase and to confirm its nuclear pore complex localization. Nuclear pores are the sites at which movement of proteins, nucleic acids, and nucleoprotein complexes take place between the cytoplasm and the nucleus. This is an important area of current research in cellular biology, since communication between the nucleus and the cytoplasm is critical to the eukaryotic cell. Very little is known about the transport process or the macromolecules which are involved. It is generally accepted that there is an ATPase associated with nuclear pores. Affinity photolabeling of a candidate nuclear pore ATPase led to the identification of the putative myosin-like pore protein, which will be studied further with the support of this award.
