Three v-SNAREs and Two t-SNAREs, Present in a Pentameric cis-SNARE Complex on Isolated Vacuoles, Are Essential for Homotypic Fusion

• DOI: 10.1083/jcb.145.7.1435
• Publication Date: 2002-07-26T16:45:41Z
• Authors: Christian Ungermann; Gabriele Fischer ...; Ole N. Jensen; Nathan Margolis; Tom H. Stevens; William Wickner
• Keywords: SNARE complex

Vacuole SNAREs, including the t-SNAREs Vam3p and Vam7p v-SNARE Nyv1p, are found in a multisubunit “cis” complex on isolated organelles. We now identify v-SNAREs Vti1p Ykt6p by mass spectrometry as additional components of immunoisolated vacuolar SNARE complex. Immunodepletion detergent extracts with anti-Vti1p removes all that is Vam3p, immunodepletion anti-Ykt6p complexed anti-Nyv1p other demonstrating they together same cis multi-SNARE After priming, which disassembles cis-SNARE complex, antibodies to any five proteins still inhibit fusion assay until docking stage completed, suggesting each plays role docking. Furthermore, vti1 temperature-sensitive alleles cause synthetic fusion-defective phenotype our reaction. Our data show vacuole-vacuole requires Vti1p, Ykt6p.