The Arabidopsis R-SNARE VAMP721 Interacts with KAT1 and KC1 K+ Channels to Moderate K+ Current at the Plasma Membrane. Issue 6 (22nd May 2015)

• Record Type: Journal Article
• Authors: Zhang, Ben; Karnik, Rucha; Wang, Yizhou; Wallmeroth, Niklas; Blatt, Michael R.; Grefen, Christopher
• Is Part Of: The Plant Cell. Volume 27:Issue 6(2015)
• Page Start: 1697
• Page End: 1717
• Publication Date: 2015-05-22
• DOI: 10.1105/tpc.15.00305
• Languages: English
• ISSNs: 1040-4651

Abstract

Abstract : Opposing actions of vesicle trafficking proteins on K + channel gating implicate a 'handoff' in binding and channel control during vesicle fusion. Abstract: SNARE (soluble N -ethylmaleimide-sensitive factor protein attachment protein receptor) proteins drive vesicle traffic, delivering membrane and cargo to target sites within the cell and at its surface. They contribute to cell homeostasis, morphogenesis, and pathogen defense. A subset of SNAREs, including the Arabidopsis thaliana SNARE SYP121, are known also to coordinate solute uptake via physical interactions with K + channels and to moderate their gating at the plasma membrane. Here, we identify a second subset of SNAREs that interact to control these K + channels, but with opposing actions on gating. We show that VAMPs (vesicle-associated membrane proteins), which target vesicles to the plasma membrane, also interact with and suppress the activities of the inward-rectifying K + channels KAT1 and KC1. Interactions were evident in yeast split-ubiquitin assays, they were recovered in vivo by ratiometric bimolecular fluorescence complementation, and they were sensitive to mutation of a single residue, Tyr-57, within the longin domain of VAMP721. Interaction was also recovered on exchange of the residue at this site in the homolog VAMP723, which normally localizes to the endoplasmic reticulum and otherwise did not interact. Functional analysis showed reduced channel activity and alterations in voltage sensitivity that are best explained by a physical interaction with the channel gates. These actions complement those of SYP121, a cognate SNARE partner of VAMP721, and lead us to propose that the channel interactions reflect a "hand-off" in channel control between the two SNARE proteins that is woven together with vesicle fusion.