A t-SNARE of the endocytic pathway must be activated for fusion

doi:10.1083/jcb.200104092

Published 10 December 2001. doi:10.1083/jcb.200104092

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© The Rockefeller University Press, 0021-9525/2001/12/961 $5.00
The Journal of Cell Biology, Volume 155, Number 6, December 10, 2001 961-968

Article

A t-SNARE of the endocytic pathway must be activated for fusion

Fabienne Paumet¹, Britta Brügger², Francesco Parlati¹, James A. McNew³, Thomas H. Söllner¹ and James E. Rothman¹

¹ Cellular Biochemistry and Biophysics Program, Memorial Sloan Kettering Cancer Center, New York, NY 10021
² Biochemie-Zentrum, Im Neuenheimer Feld 328, 69120 Heidelberg, Germany
³ Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77251

Address correspondence to James E. Rothman, Cellular Biochemistry and Biophysics Program, Memorial Sloan Kettering Cancer Center, 1275 York Ave., Box 251, New York, NY 10021. Tel.: (212) 639-8598. Fax: (212) 717-3604. E-mail: j-rothman{at}ski.mskcc.org

The t-SNARE in a late Golgi compartment (Tlg2p) syntaxin isrequired for endocytosis and localization of cycling proteinsto the late Golgi compartment in yeast. We show here that Tlg2passembles with two light chains, Tlg1p and Vti1p, to form afunctional t-SNARE that mediates fusion, specifically with thev-SNAREs Snc1p and Snc2p. In vitro, this t-SNARE is inert, lockedin a nonfunctional state, unless it is activated for fusion.Activation can be mediated by a peptide derived from the v-SNARE,which likely bypasses additional regulatory proteins in thecell. Locking t-SNAREs creates the potential for spatial andtemporal regulation of fusion by signaling processes that unleashtheir fusion capacity.

Key Words: SNARE; endosome; Tlg2p; Snc2p; fusion

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