Indication for differential sorting of the rat v-SNARE splice isoforms VAMP-1a and -1b
| Autor: | Michael Bette, Gabriel A. Bonaterra, Ralf Jacob, Susanne Wiegand, Hans-Georg Lüers, Fiona R. Rodepeter, Robert Mandic, Anson W. Lowe |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Vesicle-Associated Membrane Protein 1 Cell Biology Biology Biochemistry Rats Cell biology 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Microscopy Fluorescence Tumor Cells Cultured Animals Humans Protein Isoforms Splice isoforms Attachment protein Receptor Molecular Biology 030217 neurology & neurosurgery Intracellular |
| Zdroj: | Biochemistry and Cell Biology. 95:500-509 |
| ISSN: | 1208-6002 0829-8211 |
| DOI: | 10.1139/bcb-2016-0184 |
| Popis: | Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins are essential constituents of the intracellular trafficking machinery. The variable C-terminus in the 2 rat VAMP-1 splice isoforms VAMP-1a and -1b potentially acts as a sorting signal, because similar changes at the C-terminal end of a human VAMP-1 splice isoform resulted in its sorting to mitochondria. To evaluate the differences in the subcellular localization of these two v-SNARE proteins, VAMP-1a and -1b proteins tagged with green fluorescent protein (GFP) and red fluorescent protein (RFP) were expressed in HeLa, COS-7, and MDCK cells and evaluated by conventional confocal as well as total internal reflection fluorescence microscopy. Regions consistent with the endoplasmic reticulum and Golgi apparatus demonstrated a major overlap of both signals. In the periphery, vesicular structures were observed that mainly expressed one of the 2 isoforms. Within our experimental settings, we could not observe sorting of any of the 2 isoforms to mitochondria or peroxisomes, whereas both isoforms were found expressed in a minor subset of singular vesicles, which sporadically appeared to co-localize with the exocyst marker EXOC3/Sec6. Because vesicular structures were seen that expressed only one of the two splice variants, it is possible that VAMP-1a and VAMP-1b are sorted to distinct cellular compartments that require further characterization. |
| Databáze: | OpenAIRE |
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