Naomi Donald

Summary: Early responses of plants to environmental stress factors prevent damage but can delay growth and development in fluctuating conditions. Optimising these trade‐offs requires tunability of plant responsiveness to environmental signals. We have previously reported that Histone Deacetylase Complex 1 (HDC1), which interacts with multiple proteins in histone deacetylation complexes, regulates the stress responsiveness of Arabidopsis seedlings, but the underlying mechanism remained elusive. Here, we show that HDC1 attenuates transcriptome re‐programming in salt‐treated seedlings, and we identify two genes ( LEA and MAF5 ) that inhibit seedling establishment under salt stress downstream of HDC1. HDC1 attenuates their transcriptional induction by salt via a dual mechanism involving H3K9/14 deacetylation and H3K27 trimethylation. The latter, but not the former, was also abolished in a triple knockout mutant of the linker histone H1, which partially mimics the hypersensitivity of the hdc1‐1 mutant to salt stress. Although stress‐induced H3K27me3 accumulation required both H1 and HDC1, it was not fully recovered by complementing hdc1‐1 with a truncated, H1‐binding competent HDC1 suggesting other players or independent inputs. The combined findings reveal a dual brake function of HDC1 via regulating both active and repressive epigenetic marks on stress‐inducible genes. This natural ‘anti‐panic’ device offers a molecular leaver to tune stress responsiveness in plants.

Direct interactions between exocyst and SNARE subunits hierarchically coordinate secretion at the plasma membrane for plant growth.

Cell expansion requires that ion transport and secretory membrane traffic operate in concert. Evidence from Arabidopsis (Arabidopsis thaliana) indicates that such coordination is mediated by physical interactions between subsets of so-called SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins, which drive the final stages of vesicle fusion, and K⁺ channels, which facilitate uptake of the cation to maintain cell turgor pressure as the cell expands. However, the sequence of SNARE binding with the K⁺ channels and its interweaving within the events of SNARE complex assembly for exocytosis remains unclear. We have combined protein-protein interaction and electrophysiological analyses to resolve the binding interactions of the hetero-oligomeric associations. We find that the RYxxWE motif, located within the voltage sensor of the K⁺ channels, is a nexus for multiple SNARE interactions. Of these, K⁺ channel binding and its displacement of the regulatory protein SEC11 is critical to prime the Qa-SNARE SYP121. Our results indicate a stabilizing role for the Qbc-SNARE SNAP33 in the Qa-SNARE transition to SNARE complex assembly with the R-SNARE VAMP721. They also suggest that, on its own, the R-SNARE enters an anomalous binding mode with the channels, possibly as a fail-safe measure to ensure a correct binding sequence. Thus, we suggest that SYP121 binding to the K⁺ channels serves the role of a primary trigger to initiate assembly of the secretory machinery for exocytosis.

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.

Channel binding depends on coordination between the two binding sites on VAMP721, consistent with the K+ channels as nucleation points for SNARE complex assembl