Dual Sites for SEC11 on the SNARE SYP121 Implicate a Binding Exchange during Secretory Traffic.

SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins facilitate vesicle traffic through their assembly in a heteromeric complex that drives membrane fusion. Much of vesicle traffic at the Arabidopsis (Arabidopsis thaliana) plasma membrane is subject to the Sec1/Munc18 protein SEC11, which, along with plasma membrane K⁺ channels, selectively binds with the SNARE SYP121 to regulate its assembly in complex. How SEC11 binding is coordinated with the K⁺ channels is poorly understood, as both SEC11 and the channels are thought to compete for the same SNARE binding site. Here, we identify a second binding motif within the N terminus of SYP121 and demonstrate that this motif affects SEC11 binding independently of the F⁹xRF motif that is shared with the K⁺ channels. This second, previously unrecognized motif is centered on residues R²⁰R²¹ of SYP121 and is essential for SEC11 interaction with SYP121. Mutation of the R²⁰R²¹ motif blocked vesicle traffic without uncoupling the effects of SYP121 on solute and K⁺ uptake associated with the F⁹xRF motif; the mutation also mimicked the effects on traffic block observed on coexpression of the dominant-negative SEC11^Δ149 fragment. We conclude that the R²⁰R²¹ motif represents a secondary site of interaction for the Sec1/Munc18 protein during the transition of SYP121 from the occluded to the open conformation that leads to SNARE complex assembly.

KEYWORDS: {{ getKeywords(articleDetailText.words) }}