Distinct SNARE complexes mediating membrane fusion in Golgi transport based on combinatorial specificity Journal Article
| Authors: | Parlati, F.; Varlamov, O.; Paz, K.; Mcnew, J. A.; Hurtado, D.; Söllner, T. H.; Rothman, J. E. |
| Article Title: | Distinct SNARE complexes mediating membrane fusion in Golgi transport based on combinatorial specificity |
| Abstract: | Syntaxin-5 (Sed5) is the only syntaxin needed for transport into and across the yeast Golgi, raising the question of how a single syntaxin species could mediate vesicle transport in both the anterograde and the retrograde direction within the stack. Sed5 is known to combine with two light chains (Bosl and Sec22) to form the t-SNARE needed to receive vesicles from the endoplasmic reticulum. However, the yeast Golgi contains several other potential light chains with which Sed5 could potentially combine to form other t-SNAREs. To explore the degree of specificity in the choice of light chains by a t-SNARE, we undertook a comprehensive examination of the capacity of all 21 Sed5-based t-SNAREs that theoretically could assemble in the yeast Golgi to fuse with each of the 7 potential v-SNAREs also present in this organelle. Only one additional of these 147 combinations was fusogenic. This functional proteomic strategy thereby revealed a previously uncharacterized t-SNARE in which Sed5 is the heavy chain and Gos1 and Ykt6 are the light chains, and whose unique cognate v-SNARE is Sftl. Immunoprecipitation experiments confirmed the existence of this complex in vivo. Fusion mediated by this second Golgi SNAREpin is topologically restricted, and existing genetic and morphologic evidence implies that it is used for transport across the Golgi stack. From this study, together with the previous functional proteomic analyses which have tested 275 distinct quaternary SNARE combinations, it follows that the fusion potential and transport pathways of the yeast cell can be read out from its genome sequence according to the SNARE hypothesis with a predictive accuracy of about 99.6%. |
| Keywords: | unclassified drug; gene sequence; sequence analysis; nonhuman; conference paper; protein conformation; protein analysis; complex formation; protein assembly; protein protein interaction; protein binding; membrane proteins; intracellular transport; proteomics; time factors; endoplasmic reticulum; recombinant fusion proteins; cell membrane; peptides; glutathione transferase; immunoprecipitation; plasmids; binding site; microscopy, fluorescence; protein structure, tertiary; saccharomyces cerevisiae proteins; golgi complex; fungal proteins; vesicular transport proteins; intracellular membranes; membrane fusion; snare protein; syntaxin; syntaxin 5; qa-snare proteins; qb-snare proteins; golgi apparatus; r-snare proteins; snare proteins; organelle; precipitin tests; priority journal; qc-snare proteins; syntaxin vesicle |
| Journal Title: | Proceedings of the National Academy of Sciences of the United States of America |
| Volume: | 99 |
| Issue: | 8 |
| ISSN: | 0027-8424 |
| Publisher: | National Academy of Sciences |
| Date Published: | 2002-04-16 |
| Start Page: | 5424 |
| End Page: | 5429 |
| Language: | English |
| DOI: | 10.1073/pnas.082100899 |
| PUBMED: | 11959998 |
| PROVIDER: | scopus |
| PMCID: | PMC122785 |
| DOI/URL: | |
| Notes: | Export Date: 14 November 2014 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work