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Originally published as MBC in Press, 10.1091/mbc.E03-07-0535 on October 17, 2003

Vol. 15, Issue 1, 162-175, January 2004

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rsly1 Binding to Syntaxin 5 Is Required for Endoplasmic Reticulum-to-Golgi Transport but Does Not Promote SNARE Motif Accessibility

Antionette L. Williams, Sebastian Ehm, Noëlle C. Jacobson, Dalu Xu, and Jesse C. Hay *

University of Michigan, Department of Molecular, Cellular, and Developmental Biology, Ann Arbor, Michigan 48109-1048

Submitted July 26, 2003; Revised September 8, 2003; Accepted September 16, 2003
Monitoring Editor: Keith Mostov

Although some of the principles of N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) function are well understood, remarkably little detail is known about sec1/munc18 (SM) protein function and its relationship to SNAREs. Popular models of SM protein function hold that these proteins promote or maintain an open and/or monomeric pool of syntaxin molecules available for SNARE complex formation. To address the functional relationship of the mammalian endoplasmic reticulum/Golgi SM protein rsly1 and its SNARE binding partner syntaxin 5, we produced a conformation-specific monoclonal antibody that binds only the available, but not the cis-SNARE–complexed nor intramolecularly closed form of syntaxin 5. Immunostaining experiments demonstrated that syntaxin 5 SNARE motif availability is nonuniformly distributed and focally regulated. In vitro endoplasmic reticulum-to-Golgi transport assays revealed that rsly1 was acutely required for transport, and that binding to syntaxin 5 was absolutely required for its function. Finally, manipulation of rsly1–syntaxin 5 interactions in vivo revealed that they had remarkably little impact on the pool of available syntaxin 5 SNARE motif. Our results argue that although rsly1 does not seem to regulate the availability of syntaxin 5, its function is intimately associated with syntaxin binding, perhaps promoting a later step in SNARE complex formation or function.


Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E03–07–0535. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E03-07-0535.

* Corresponding author. E-mail address: jessehay{at}umich.edu.




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