A multigene family encoding R-SNAREs in the ciliate Paramecium tetraurelia.
SNARE proteins (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) mediate membrane interactions and are conventionally divided into Q-SNAREs and R-SNAREs according to the possession of a glutamine or arginine residue at the core of their SNARE domain. Here, we describe a set of R-SNAREs from the ciliate Paramecium tetraurelia consisting of seven families encoded by 12 genes that are expressed simultaneously. The complexity of the endomembrane system in Paramecium can explain this high number of genes. All P. tetraurelia synaptobrevins (PtSybs) possess a SNARE domain and show homology to the Longin family of R-SNAREs such as Ykt6, Sec22 and tetanus toxin-insensitive VAMP (TI-VAMP). We localized four exemplary PtSyb subfamilies with GFP constructs and antibodies on the light and electron microscopic level. PtSyb1-1, PtSyb1-2 and PtSyb3-1 were found in the endoplasmic reticulum, whereas PtSyb2 is localized exclusively in the contractile vacuole complex. PtSyb6 was found cytosolic but also resides in regularly arranged structures at the cell cortex (parasomal sacs), the cytoproct and oral apparatus, probably representing endocytotic compartments. With gene silencing, we showed that the R-SNARE of the contractile vacuole complex, PtSyb2, functions to maintain structural integrity as well as functionality of the osmoregulatory system but also affects cell division.
Chair of Cell Biology and Ultrastructure Research, University of Konstanz, PO Box 5560, 78457 Konstanz, Germany. christina.schilde@uni-konstanzde
Christina
C
Schilde
Thomas
T
Wassmer
Joerg
J
Mansfeld
Helmut
H
Plattner
Roland
R
Kissmehl
D000595:Amino Acid Sequence; D000818:Animals; D002458:Cell Fractionation; D048430:Cell Shape; D004721:Endoplasmic Reticulum / Q000378:metabolism / Q000648:ultrastructure; D020868:Gene Silencing; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D005810:Multigene Family; D016805:Paramecium tetraurelia / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D015800:Protozoan Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism; D050683:R-SNARE Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
D015800:Protozoan Proteins; D050683:R-SNARE Proteins; D011993:Recombinant Fusion Proteins