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THE PLANT CELL, Vol 9, Issue 2 157-169, Copyright © 1997 by American Society of Plant Biologists
Dynamics of Phragmoplastin in Living Cells during Cell Plate Formation and Uncoupling of Cell Elongation from the Plane of Cell Division
X. Gu and DPS. Verma
Department of Molecular Genetics, Ohio State Biochemistry Program and Plant Biotechnology Center, Ohio State University, Columbus, Ohio 43210
The cell plate is formed by the fusion of Golgi apparatus-derived vesicles
in the center of the phragmoplast during cytokinesis in plant cells. A
dynamin-like protein, phragmoplastin, has been isolated and shown to be
associated with cell plate formation in soybean by using
immunocytochemistry. In this article, we demonstrate that similar to
dynamin, phragmoplastin polymerizes to form oligomers. We fused soybean
phragmoplastin with the green fluorescence protein (GFP) and introduced it
into tobacco BY-2 cells to monitor the dynamics of early events in cell
plate formation. We demonstrate that the chimeric protein is functional and
targeted to the cell plate during cytokinesis in transgenic cells.
GFP-phragmoplastin was found to appear first in the center of the forming
cell plate, and as the cell plate grew outward, it redistributed to the
growing margins of the cell plate. The redistribution of phragmoplastin may
require microtubule reorganization because the microtubule-stabilizing drug
taxol inhibited phragmoplastin redistribution. Our data show that
throughout the entire process of cytokinesis, phragmoplastin is
concentrated in the area in which membrane fusion is active, suggesting
that phragmoplastin participates in an early membrane fusion event during
cell plate formation. Based on the dynamics of GFP-phragmoplastin, it
appears that the process of cell plate formation is completed in two
phases. The first phase is confined to the cylinder of the phragmoplast
proper and is followed by a second phase that deposits phragmoplast
vesicles in a concentric fashion, resulting in a ring of fluorescence, with
the concentration of vesicles being higher at the periphery. In addition,
overexpression of GFP-phragmoplastin appears to act as a dominant negative,
slowing down the completion of cell plate formation, and often results in
an oblique cell plate. The latter appears to uncouple cell elongation from
the plane of cell division, forming twisted and elongated cells with
longitudinal cell divisions.
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