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The  D. melanogaster germ cell-less (gcl)  gene has previously been shown to play a key role in the establishment of the germ cell lineage during fly embryogenesis. To identify other molecules that function with Gcl in this process, we have conducted a yeast two-hybrid screen that utilized Gcl protein as bait. A predominant class of Gcl-interacting clones encodes a species of importin-α from  Drosophila (karyopherin-α3; kap-α3) , a nuclear-localization sequence binding protein previously shown to act in the transport of proteins from the cytoplasm to the nucleus. The expression of  kap-α3  is widespread both temporally and spatially throughout the embryo during development, as judged by Northern blotting and whole-mount  in situ  hybridization to  Drosophila  embryos, suggesting that it functions at multiple stages of development. Studies of the Gcl/Kap-α3 interaction have identified a functional nuclear- localization sequence in Gcl protein which is necessary for an  in vivo  interaction and for nuclear entry of Gcl, making it likely that one role for Kap-α3 is to deliver Gcl protein to the nucleus. The identification of Kap-α3 and an  in vivo  substrate will allow for further characterization of the basis for specificity between importin-αmolecules and their binding substrates.

Cloning of karyopherin-α3 from Drosophila Through Its Interaction with the Nuclear Localization Sequence of Germ Cell-Less Protein