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© The Rockefeller University Press, 0021-9525/1997//1485 $5.00
The Journal of Cell Biology, Volume 139, Number 6, , 1997 1485-1493

The Arabidopsis KNOLLE Protein Is a Cytokinesis-specific Syntaxin

Martina H. Lauber^*, Irene Waizenegger^*, Thomas Steinmann^*, Heinz Schwarz, Ulrike Mayer^*, Inwhan Hwang, Wolfgang Lukowitz^*, and Gerd Jürgens^*

^* Lehrstuhl für Entwicklungsgenetik, Universität Tübingen, D-72076 Tübingen, Federal Republic of Germany; Max-Planck-Institut für Entwicklungsbiologie, D-72076 Tübingen, Federal Republic of Germany; and Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Chinju, 660-701, Korea

In higher plant cytokinesis, plasma membrane and cell wall originate by vesicle fusion in the plane of cell division. The Arabidopsis KNOLLE gene, which is required for cytokinesis, encodes a protein related to vesicle-docking syntaxins. We have raised specific rabbit antiserum against purified recombinant KNOLLE protein to show biochemically and by immunoelectron microscopy that KNOLLE protein is membrane associated. Using immunofluorescence microscopy, KNOLLE protein was found to be specifically expressed during mitosis and, unlike the plasma membrane H⁺-ATPase, to localize to the plane of division during cytokinesis. Arabidopsis dynamin-like protein ADL1 accumulates at the plane of cell plate formation in knolle mutant cells as in wild-type cells, suggesting that cytokinetic vesicle traffic is not affected. Furthermore, electron microscopic analysis indicates that vesicle fusion is impaired. KNOLLE protein was detected in mitotically dividing cells of various parts of the developing plant, including seedling root, inflorescence meristem, floral meristems and ovules, and the cellularizing endosperm, but not during cytokinesis after the male second meiotic division. Thus, KNOLLE is the first syntaxin-like protein that appears to be involved specifically in cytokinetic vesicle fusion.

Abbreviations used in this paper: ADL1, Arabidopsis dynamin-like protein; DAPI, 4',6-diamidino-2-phenylindole; KN, KNOLLE; MTSB, microtubule-stabilizing buffer; PM ATPase, plasma membrane H⁺-ATPase.

Address all correspondence to Gerd Jürgens Lehrstuhl für Entwicklungsgenetik, Universität Tübingen, Spemannstr. 37-39, D-72076 Tübingen, Federal Republic of Germany. Tel.: 49-7071-601-861. Fax: 49-7071-601-862. E-mail: geju{at}mpib-tuebingen.mpg.de

Irene Waizenegger's present address is Institute of Molecular Pathology, A-1030, Vienna, Austria.

Wolfgang Lukowitz's present address is Department of Plant Biology, Carnegie Institution of Washington, Stanford, CA 94305.

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