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© The Rockefeller University Press, 0021-9525/1997//291 $5.00
The Journal of Cell Biology, Volume 137, Number 2, , 1997 291-303

Article

The Dynamic Nuclear Redistribution of an hnRNP K-homologous Protein during Drosophila Embryo Development and Heat Shock. Flexibility of Transcription Sites In Vivo



Peter Buchenau*, Harald Saumweber||, and Donna J. Arndt-Jovin*

* Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, 370170 Göttingen, Germany; and || Institute of Biology, Department of Cytogenetics, Humboldt University of Berlin, 10115 Berlin, Germany

The Drosophila protein Hrb57A has sequence homology to mammalian heterogenous nuclear ribonucleoprotein (hnRNP) K proteins. Its in vivo distribution has been studied at high resolution by confocal laser scanning microscopy (CLSM) in embryos injected with fluorescently labeled monoclonal antibody. Injection of antibody into living embryos had no apparent deleterious effects on further development. Furthermore, the antibody-protein complex could be observed for more than 7 cell cycles in vivo, revealing a dynamic redistribution from the nucleus to cytoplasm at each mitosis from blastoderm until hatching. The evaluation of two- and three-dimensional CLSM data sets demonstrated important differences in the localization of the protein in the nuclei of living compared to fixed embryos. The Hrb57A protein was recruited to the 93D locus upon heat shock and thus serves as an in vivo probe for the activity of the gene in diploid cells of the embryo. Observations during heat shock revealed considerable mobility within interphase nuclei of this transcription site. Furthermore, the reinitiation as well as the down regulation of transcriptional loci in vivo during the recovery from heat shock could be followed by the rapid redistribution of the hnRNP K during stress recovery. These data are incompatible with a model of the interphase nucleus in which transcription complexes are associated with a rigid nuclear matrix.

Abbreviations used in this paper: CLSM, confocal laser scanning microscopy; hn, heterogeneous nuclear; 3-D, three dimensional; sn, small nuclear.

Please address all correspondence to Donna Arndt-Jovin, Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, 370170 Gottingen, Germany. Tel.: (49) 551-2011-393; Fax.: (49) 551-2011467; E-mail: djovin{at}mpc186.mpibpc.gwdg.de

P. Buchenau's present address is Botanical Institute, University of Bonn, 53115 Bonn, Germany.


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