Distinct SNARE complexes mediating membrane fusion in Golgi transport based on combinatorial specificity

Francesco Parlati, Oleg Varlamov, Keren Paz, James A. McNew, David Hurtado, Thomas H. Söllner, James E. Rothman

Research output: Contribution to journalArticle

130 Citations (Scopus)

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%.

Original languageEnglish (US)
Pages (from-to)5424-5429
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume99
Issue number8
DOIs
StatePublished - Apr 16 2002
Externally publishedYes

Fingerprint

SNARE Proteins
Membrane Fusion
Qa-SNARE Proteins
Yeasts
Light
Proteomics
Transport Vesicles
Immunoprecipitation
Endoplasmic Reticulum
Organelles
Genome

Keywords

  • Organelle
  • Syntaxin vesicle

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Distinct SNARE complexes mediating membrane fusion in Golgi transport based on combinatorial specificity. / Parlati, Francesco; Varlamov, Oleg; Paz, Keren; McNew, James A.; Hurtado, David; Söllner, Thomas H.; Rothman, James E.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 99, No. 8, 16.04.2002, p. 5424-5429.

Research output: Contribution to journalArticle

Parlati, Francesco ; Varlamov, Oleg ; Paz, Keren ; McNew, James A. ; Hurtado, David ; Söllner, Thomas H. ; Rothman, James E. / Distinct SNARE complexes mediating membrane fusion in Golgi transport based on combinatorial specificity. In: Proceedings of the National Academy of Sciences of the United States of America. 2002 ; Vol. 99, No. 8. pp. 5424-5429.
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