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Eukaryotic Cell, August 2008, p. 1387-1402, Vol. 7, No. 8
1535-9778/08/$08.00+0     doi:10.1128/EC.00012-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Molecular Identification of a SNAP-25-Like SNARE Protein in Paramecium{triangledown}

Christina Schilde,* Kaya Lutter,{dagger} Roland Kissmehl, and Helmut Plattner

Department of Biology, University of Konstanz, 78457 Konstanz, Germany

Received 8 January 2008/ Accepted 25 May 2008

Using database searches of the completed Paramecium tetraurelia macronuclear genome with the metazoan SNAP-25 homologues, we identified a single 21-kDa Qb/c-SNARE in this ciliated protozoan, named P. tetraurelia SNAP (PtSNAP), containing the characteristic dual heptad repeat SNARE motifs of SNAP-25. The presence of only a single Qb/c class SNARE in P. tetraurelia is surprising in view of the multiple genome duplications and the high number of SNAREs found in other classes of this organism. As inferred from the subcellular localization of a green fluorescent protein (GFP) fusion construct, the protein is localized on a variety of intracellular membranes, and there is a large soluble pool of PtSNAP. Similarly, the PtSNAP that is detected with a specific antibody in fixed cells is associated with a number of intracellular membrane structures, including food vacuoles, the contractile vacuole system, and the sites of constitutive endo- and exocytosis. Surprisingly, using gene silencing, we could not assign a role to PtSNAP in the stimulated exocytosis of dense core vesicles (trichocysts), but we found an increased number of food vacuoles in PtSNAP-silenced cells. In conclusion, we identify PtSNAP as a Paramecium homologue of metazoan SNAP-25 that shows several divergent features, like resistance to cleavage by botulinum neurotoxins.

* Corresponding author. Mailing address: Department of Biology, University of Konstanz, P.O. Box 5560, 78457 Konstanz, Germany. Phone: 49-7531-88-4230. Fax: 49-7531-88-2245. E-mail: christina.schilde{at}uni-konstanz.de

{triangledown} Published ahead of print on 13 June 2008.

{dagger} Present address: Institut für Biochemie und Molekularbiologie I, Universitätsklinikum Düsseldorf, Moorenstrasse 5, 40225 Düsseldorf, Germany.

Eukaryotic Cell, August 2008, p. 1387-1402, Vol. 7, No. 8
1535-9778/08/$08.00+0     doi:10.1128/EC.00012-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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