Molecular dissection of plant cytokinesis and phragmoplast structure: a survey of GFP-tagged proteins

Daniël Van Damme, François-Yves Bouget¹, Kris Van Poucke, Dirk Inzé², and Danny Geelen

Department of Plant Systems Biology, Flanders Interuniversity Institute for Biotechnology, Ghent University, Technologiepark 927, B-9052 Gent, Belgium

¹ current address: Laboratoire Arago, UMR7628 CNRS, Université Pierre et Marie Curie, BP 44, 66651 Banyuls sur Mer cedex, France  

² to whom correspondence should be adressed. E-mail dirk.inze@psb.ugent.be; fax 32-9-3313809

Abstract

To identify molecular players implicated in cytokinesis and division plane determination, the Arabidopsis thaliana genome was explored for potential cytokinesis genes. More than 100 open reading frames were selected based on similarity to yeast and animal cytokinesis genes, cytoskeleton and polarity genes, and Nicotiana tabacum genes showing cell cycle controlled expression. The subcellular localization of these proteins was determined by means of GFP-tagging in tobacco Bright Yellow-2 (BY-2) cells and Arabidopsis plants. Detailed confocal microscopy identified 15 proteins targeted to distinct regions of the phragmoplast and the cell plate. EB1- and MAP65-like proteins were associated with the plus-end, the minus-end, or along the entire length of microtubules. The actin-binding protein myosin, the kinase Aurora, and a novel cell cycle protein designated T22, accumulated preferentially at the midline. EB1 and Aurora, in addition to other regulatory proteins (homologs of Mob1, Sid1, and Sid2), were targeted to the nucleus, suggesting that this organelle operates as a coordinating hub for cytokinesis.

Links to supplemental material:

all localisations in the database are C-terminal fusion proteins with EGFP, 

expressed under the 35S promotor unless otherwise listed.

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