Binding of SEC 11 Indicates Its Role in SNARE Recycling after Vesicle Fusion and Identi fi es Two Pathways for Vesicular Traf fi c to the Plasma Membrane

SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins drive vesicle fusion in all eukaryotes and contribute to homeostasis, pathogen defense, cell expansion, and growth in plants. Two homologous SNAREs, SYP121 (=SYR1/PEN1) and SYP122, dominate secretory traffic to the Arabidopsis thaliana plasma membrane. Although these proteins overlap functionally, differences between SYP121 and SYP122 have surfaced, suggesting that they mark two discrete pathways for vesicular traffic. The SNAREs share primary cognate partners, which has made separating their respective control mechanisms difficult. Here, we show that the regulatory protein SEC11 (=KEULE) binds selectively with SYP121 to affect secretory traffic mediated by this SNARE. SEC11 rescued traffic block by dominant-negative (inhibitory) fragments of both SNAREs, but only in plants expressing the native SYP121. Traffic and its rescue were sensitive to mutations affecting SEC11 interaction with the N terminus of SYP121. Furthermore, the domain of SEC11 that bound the SYP121 N terminus was itself able to block secretory traffic in the wild type and syp122 but not in syp121 mutant Arabidopsis. Thus, SEC11 binds and selectively regulates secretory traffic mediated by SYP121 and is important for recycling of the SNARE and its cognate partners.

[1]  Jennifer L. Martin,et al.  SNARE-ing the structures of Sec1/Munc18 proteins. , 2014, Current opinion in structural biology.

[2]  D. Fasshauer,et al.  The SM protein Sly1 accelerates assembly of the ER–Golgi SNARE complex , 2014, Proceedings of the National Academy of Sciences.

[3]  G. De Jaeger,et al.  The Phragmoplast-Orienting Kinesin-12 Class Proteins Translate the Positional Information of the Preprophase Band to Establish the Cortical Division Zone in Arabidopsis thaliana[C][W] , 2014, Plant Cell.

[4]  The split-ubiquitin system for the analysis of three-component interactions. , 2014, Methods in molecular biology.

[5]  M. Blatt,et al.  Applications of fluorescent marker proteins in plant cell biology. , 2014, Methods in molecular biology.

[6]  P. Schulze-Lefert,et al.  Requirement of vesicle-associated membrane protein 721 and 722 for sustained growth during immune responses in Arabidopsis , 2013, Molecules and cells.

[7]  Abhijit Karnik,et al.  SDM-Assist software to design site-directed mutagenesis primers introducing “silent” restriction sites , 2013, BMC Bioinformatics.

[8]  T. Südhof,et al.  The membrane fusion enigma: SNAREs, Sec1/Munc18 proteins, and their accomplices--guilty as charged? , 2012, Annual review of cell and developmental biology.

[9]  Christopher Grefen,et al.  A 2in1 cloning system enables ratiometric bimolecular fluorescence complementation (rBiFC). , 2012, BioTechniques.

[10]  Adrien S. Chevalier,et al.  Selective Regulation of Maize Plasma Membrane Aquaporin Trafficking and Activity by the SNARE SYP121[W] , 2012, Plant Cell.

[11]  D. James,et al.  Low-resolution solution structures of Munc18:Syntaxin protein complexes indicate an open binding mode driven by the Syntaxin N-peptide , 2012, Proceedings of the National Academy of Sciences.

[12]  G. Jürgens,et al.  Sec1/Munc18 protein stabilizes fusion-competent syntaxin for membrane fusion in Arabidopsis cytokinesis. , 2012, Developmental cell.

[13]  M. Blatt,et al.  Do Calcineurin B-Like Proteins Interact Independently of the Serine Threonine Kinase CIPK23 with the K+ Channel AKT1? Lessons Learned from a Ménage à Trois , 2012, Plant Physiology.

[14]  M. Blatt,et al.  The trafficking protein SYP121 of Arabidopsis connects programmed stomatal closure and K⁺ channel activity with vegetative growth. , 2012, The Plant journal : for cell and molecular biology.

[15]  G. Jürgens,et al.  Mechanisms of Functional Specificity Among Plasma‐Membrane Syntaxins in Arabidopsis , 2011, Traffic.

[16]  M. Blatt,et al.  Ion transport, membrane traffic and cellular volume control. , 2011, Current opinion in plant biology.

[17]  M. Blatt,et al.  A molecular framework for coupling cellular volume and osmotic solute transport control. , 2011, Journal of experimental botany.

[18]  Jennifer L. Martin,et al.  Possible roles for Munc18-1 domain 3a and Syntaxin1 N-peptide and C-terminal anchor in SNARE complex formation , 2010, Proceedings of the National Academy of Sciences.

[19]  E. Jorgensen,et al.  Syntaxin N-terminal peptide motif is an initiation factor for the assembly of the SNARE–Sec1/Munc18 membrane fusion complex , 2010, Proceedings of the National Academy of Sciences.

[20]  J. Kudla,et al.  A ubiquitin-10 promoter-based vector set for fluorescent protein tagging facilitates temporal stability and native protein distribution in transient and stable expression studies. , 2010, The Plant journal : for cell and molecular biology.

[21]  A. Hills,et al.  A Novel Motif Essential for SNARE Interaction with the K+ Channel KC1 and Channel Gating in Arabidopsis[W] , 2010, Plant Cell.

[22]  M. Blatt,et al.  A Tripartite SNARE-K+ Channel Complex Mediates in Channel-Dependent K+ Nutrition in Arabidopsis[W] , 2009, The Plant Cell Online.

[23]  Nia J. Bryant,et al.  The N-terminal peptide of the syntaxin Tlg2p modulates binding of its closed conformation to Vps45p , 2009, Proceedings of the National Academy of Sciences.

[24]  U. Schurr,et al.  Environmental effects on spatial and temporal patterns of leaf and root growth. , 2009, Annual review of plant biology.

[25]  A. Nakano,et al.  Differential expression control and polarized distribution of plasma membrane-resident SYP1 SNAREs in Arabidopsis thaliana. , 2009, Plant & cell physiology.

[26]  T. Südhof,et al.  Membrane Fusion: Grappling with SNARE and SM Proteins , 2009, Science.

[27]  Klaus Harter,et al.  The determination of protein-protein interactions by the mating-based split-ubiquitin system (mbSUS). , 2009, Methods in molecular biology.

[28]  M. Blatt,et al.  SNAREs--molecular governors in signalling and development. , 2008, Current opinion in plant biology.

[29]  P. Schulze-Lefert,et al.  Activity Determinants and Functional Specialization of Arabidopsis PEN1 Syntaxin in Innate Immunity* , 2008, Journal of Biological Chemistry.

[30]  M. Blatt,et al.  SNAREs: Cogs and Coordinators in Signaling and Development1 , 2008, Plant Physiology.

[31]  G. Jürgens,et al.  Co-option of a default secretory pathway for plant immune responses , 2008, Nature.

[32]  G. Dalessandro,et al.  Tomato Rab 11 a Characterization Evidenced a Difference Between SYP 121-Dependent and SYP 122-Dependent exocytosis , 2008 .

[33]  Josep Rizo,et al.  Dual Modes of Munc18-1/SNARE Interactions Are Coupled by Functionally Critical Binding to Syntaxin-1 N Terminus , 2007, The Journal of Neuroscience.

[34]  R. Panstruga,et al.  SNARE-ware: the role of SNARE-domain proteins in plant biology. , 2007, Annual review of cell and developmental biology.

[35]  Christopher Grefen,et al.  Split‐Ubiquitin System for Identifying Protein‐Protein Interactions in Membrane and Full‐Length Proteins , 2007, Current protocols in neuroscience.

[36]  Thorsten Lang,et al.  Anatomy and Dynamics of a Supramolecular Membrane Protein Cluster , 2007, Science.

[37]  M. Blatt,et al.  Abscisic Acid Triggers the Endocytosis of the Arabidopsis KAT1 K+ Channel and Its Recycling to the Plasma Membrane , 2007, Current Biology.

[38]  M. Blatt,et al.  Selective targeting of plasma membrane and tonoplast traffic by inhibitory (dominant-negative) SNARE fragments. , 2007, The Plant journal : for cell and molecular biology.

[39]  K. Findlay,et al.  The syntaxin SYP132 contributes to plant resistance against bacteria and secretion of pathogenesis-related protein 1 , 2007, Proceedings of the National Academy of Sciences.

[40]  M. Peter,et al.  Yeast split-ubiquitin-based cytosolic screening system to detect interactions between transcriptionally active proteins. , 2007, BioTechniques.

[41]  Alan Morgan,et al.  Membrane Trafficking: Three Steps to Fusion , 2007, Current Biology.

[42]  T. Südhof,et al.  Munc18-1 binds directly to the neuronal SNARE complex , 2007, Proceedings of the National Academy of Sciences.

[43]  M. Blatt,et al.  Membrane trafficking and polar growth in root hairs and pollen tubes. , 2006, Journal of experimental botany.

[44]  M. Newman,et al.  A SNARE-protein has opposing functions in penetration resistance and defence signalling pathways. , 2007, The Plant journal : for cell and molecular biology.

[45]  Reinhard Jahn,et al.  SNAREs — engines for membrane fusion , 2006, Nature Reviews Molecular Cell Biology.

[46]  M. Blatt,et al.  Selective Mobility and Sensitivity to SNAREs Is Exhibited by the Arabidopsis KAT1 K+ Channel at the Plasma Membrane[W] , 2006, The Plant Cell Online.

[47]  Heather Youngs,et al.  The PEN1 syntaxin defines a novel cellular compartment upon fungal attack and is required for the timely assembly of papillae. , 2004, Molecular biology of the cell.

[48]  B. André,et al.  K+ channel interactions detected by a genetic system optimized for systematic studies of membrane protein interactions. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[49]  M. Blatt,et al.  A new catch in the SNARE. , 2004, Trends in plant science.

[50]  A. Nakano,et al.  Systematic analysis of SNARE molecules in Arabidopsis: dissection of the post-Golgi network in plant cells. , 2004, Cell structure and function.

[51]  Keith A Mott,et al.  Changes in Surface Area of Intact Guard Cells Are Correlated with Membrane Internalization1 , 2003, Plant Physiology.

[52]  Frank Küttner,et al.  Syntaxin specificity of cytokinesis in Arabidopsis , 2003, Nature Cell Biology.

[53]  W. Weissenhorn,et al.  Structural basis for the Golgi membrane recruitment of Sly1p by Sed5p , 2002, The EMBO journal.

[54]  M. Blatt,et al.  The abscisic acid-related SNARE homolog NtSyr1 contributes to secretion and growth: evidence from competition with its cytosolic domain. , 2002, The Plant cell.

[55]  K. Misura,et al.  Self-association of the H3 Region of Syntaxin 1A , 2001, The Journal of Biological Chemistry.

[56]  Andrew A. Peden,et al.  A genomic perspective on membrane compartment organization , 2001, Nature.

[57]  G. Jürgens,et al.  The Cytokinesis Gene KEULE Encodes a Sec1 Protein That Binds the Syntaxin Knolle , 2001, The Journal of cell biology.

[58]  IMPLICATIONS FOR INTERMEDIATES IN SNARE COMPLEX ASSEMBLY , 2001 .

[59]  N. Raikhel,et al.  The Arabidopsis genome. An abundance of soluble N-ethylmaleimide-sensitive factor adaptor protein receptors. , 2000, Plant physiology.

[60]  I. Waizenegger,et al.  The Arabidopsis KNOLLE and KEULE genes interact to promote vesicle fusion during cytokinesis , 2000, Current Biology.

[61]  Richard H. Scheller,et al.  Three-dimensional structure of the neuronal-Sec1–syntaxin 1a complex , 2000, Nature.

[62]  M. Blatt.  Cellular signaling and volume control in stomatal movements in plants. , 2000, Annual review of cell and developmental biology.

[63]  M. Blatt,et al.  A tobacco syntaxin with a role in hormonal control of guard cell ion channels. , 1999, Science.

[64]  A. Brunger,et al.  Conserved structural features of the synaptic fusion complex: SNARE proteins reclassified as Q- and R-SNAREs. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[65]  S. Clough,et al.  Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[66]  I. Waizenegger,et al.  The Arabidopsis KNOLLE Protein Is a Cytokinesis-specific Syntaxin , 1997, The Journal of cell biology.

[67]  N. Chua,et al.  A glucocorticoid-mediated transcriptional induction system in transgenic plants. , 1997, The Plant journal : for cell and molecular biology.