Dual sites for SEC11 on the SNARE SYP121 implicate a binding exchange during secretory traffic

Zhang, B. , Karnik, R. , Alvim, J., Donald, N. and Blatt, M. R. (2019) Dual sites for SEC11 on the SNARE SYP121 implicate a binding exchange during secretory traffic. Plant Physiology, 180, pp. 228-239. (doi: 10.1104/pp.18.01315) (PMID:30850468) (PMCID:PMC6501095)

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Abstract

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 SEC11Δ149 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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Blatt, Professor Michael and Alvim, Mr Jonas and Karnik, Dr Rucha and Zhang, Dr Ben and Donald, Miss Naomi
Authors: Zhang, B., Karnik, R., Alvim, J., Donald, N., and Blatt, M. R.
College/School:College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
Journal Name:Plant Physiology
Publisher:American Society of Plant Biologists
ISSN:0032-0889
ISSN (Online):1532-2548
Published Online:08 March 2019
Copyright Holders:Copyright © 2019 American Society of Plant Biologists
First Published:First published in Plant Physiology 180:228-239
Publisher Policy:Reproduced under a Creative Commons License
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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
563751MAGIC - A Multi-tiered approach to generating increased carbon dioxide in the chloroplastMichael BlattBiotechnology and Biological Sciences Research Council (BBSRC)BB/I024496/1RI MOLECULAR CELL & SYSTEMS BIOLOGY
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656891Analysing GORK clustering for enhanced stomatal controlMichael BlattBiotechnology and Biological Sciences Research Council (BBSRC)BB/M001601/1RI MOLECULAR CELL & SYSTEMS BIOLOGY
650621Developing a synthetic approach to manipulating guard cell membrane transport and stomatal controlMichael BlattBiotechnology and Biological Sciences Research Council (BBSRC)BB/L019205/1RI MOLECULAR CELL & SYSTEMS BIOLOGY