Thomas C. Dockendorff, Zhenyu Tang, Thomas A. Jongens
Cloning of karyopherin-?3 from Drosophila Through
Its Interaction with the Nuclear Localization Sequence of
Germ Cell-Less Protein
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-? from Drosophila (karyopherin-?3; kap-?3),
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-?3 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-?3 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-?3 is
to deliver Gcl protein to the nucleus. The identification
of Kap-?3 and an in vivo substrate will allow for further
characterization of the basis for specificity between
importin-?molecules and their binding substrates.
Biological Chemistry, Walter de Gruyter
Print ISSN: 1431-6730
Volume: 380, 11/1999
Pages: 1263 - 1272
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