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

Cloning of karyopherin-α3 from Drosophila Through Its Interaction with the Nuclear Localization... Abstract 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biological Chemistry de Gruyter

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

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Publisher
de Gruyter
Copyright
Copyright © 1999 by the
ISSN
1431-6730
DOI
10.1515/BC.1999.161
pmid
10614818
Publisher site
See Article on Publisher Site

Abstract

Abstract 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.

Journal

Biological Chemistryde Gruyter

Published: Nov 17, 1999

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