Minimal membrane docking requirements revealed by reconstitution of Rab GTPase-dependent membrane fusion from purified components
Rab GTPases and their effectors mediate docking, the initial contact of intracellular membranes preceding bilayer fusion. However, it has been unclear whether Rab proteins and effectors are sufficient for intermembrane interactions. We have recently reported reconstituted membrane fusion that requir...
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Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 106; no. 42; pp. 17626 - 17633 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
United States
National Academy of Sciences
20.10.2009
National Acad Sciences |
Series | From the Cover: Feature Article |
Subjects | |
Online Access | Get full text |
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Summary: | Rab GTPases and their effectors mediate docking, the initial contact of intracellular membranes preceding bilayer fusion. However, it has been unclear whether Rab proteins and effectors are sufficient for intermembrane interactions. We have recently reported reconstituted membrane fusion that requires yeast vacuolar SNAREs, lipids, and the homotypic fusion and vacuole protein sorting (HOPS)/class C Vps complex, an effector and guanine nucleotide exchange factor for the yeast vacuolar Rab GTPase Ypt7p. We now report reconstitution of lysis-free membrane fusion that requires purified GTP-bound Ypt7p, HOPS complex, vacuolar SNAREs, ATP hydrolysis, and the SNARE disassembly catalysts Sec17p and Sec18p. We use this reconstituted system to show that SNAREs and Sec17p/Sec18p, and Ypt7p and the HOPS complex, are required for stable intermembrane interactions and that the three vacuolar Q-SNAREs are sufficient for these interactions. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 1C.S. and C.M.H. contributed equally to this work. 2Present address: Institute for Protein Research, Osaka University, Suita, Osaka, Japan. Author contributions: C.S., C.M.H., J.M., A.S.B., and W.W. designed research; C.S., C.M.H., J.M., and A.S.B. performed research; C.S., C.M.H., J.M., and A.S.B. contributed new reagents/analytic tools; C.S. and W.W. analyzed data; and C.S. and W.W. wrote the paper. Edited by Thomas C. Südhof, Stanford University School of Medicine, Palo Alto, CA, and approved September 16, 2009 |
ISSN: | 0027-8424 1091-6490 |
DOI: | 10.1073/pnas.0903801106 |