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Vol. 11, Issue 8, 2733-2741, August 2000
*Department of Plant Sciences, University of Arizona, Tucson,
Arizona 85721; The nucleus is a definitive feature of eukaryotic cells, comprising
twin bilamellar membranes, the inner and outer nuclear membranes, which
separate the nucleoplasmic and cytoplasmic compartments. Nuclear pores,
complex macromolecular assemblies that connect the two membranes,
mediate communication between these compartments. To explore the
morphology, topology, and dynamics of nuclei within living plant cells,
we have developed a novel method of confocal laser scanning
fluorescence microscopy under time-lapse conditions. This is used for
the examination of the transgenic expression in Arabidopsis
thaliana of a chimeric protein, comprising the GFP
(Green-Fluorescent Protein of Aequorea victoria)
translationally fused to an effective nuclear localization signal (NLS)
and to Department of Genetics and Molecular
Biology, Masaryk University, Brno, Czech Republic;
Institute of Plant Molecular Biology, Ceske Budejovice,
Czech Republic; and §Department of Genetics and Plant
Breeding, University of Technology and Agriculture, Bydgoszcz, Poland
-glucuronidase (GUS) from E. coli. This large
protein is targeted to the nucleus and accumulates exclusively within
the nucleoplasm. This article provides online access to movies
that illustrate the remarkable and unusual properties displayed by the
nuclei, including polymorphic shape changes and rapid, long-distance, intracellular movement. Movement is mediated by actin but not by
tubulin; it therefore appears distinct from mechanisms of nuclear positioning and migration that have been reported for eukaryotes. The
GFP-based assay is simple and of general applicability. It will be
interesting to establish whether the novel type of dynamic behavior
reported here, for higher plants, is observed in other eukaryotic organisms.
Online version of this article contains video material
for figures 1, 2, 3, 4, 5, and 7. Online version is available at
www.molbiolcell.org.
Present
address: Department of Biological Sciences, Stanford University,
Stanford, CA 94305.
¶
Corresponding
author. E-mail address: galbraith{at}arizona.edu.
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