Tri-SUS: a yeast split-ubiquitin assay to examine protein interactions governed by a third binding partner

Zhang, Ben; Xia, Lingfeng; Zhang, Yaxian; Wang, Hui; Karnik, Rucha

doi:10.1093/plphys/kiaa039

Cited by 2 publications

(1 citation statement)

References 14 publications

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“…How might the SNARE coordinate the association with both aquaporins and plasma membrane H + ‐ATPases? One clue comes from additional yeast Tri‐SUS analysis (Zhang et al., 2021) which showed that PIP2;1 and H + ‐ATPase AHA1 are likely to occur in a complex with the SNARE SYP132 (Figure S4). The Tri‐SUS system detects tripartite bait and prey interactions involving a third methionine‐repressible partner protein (Figure S4a).…”

Section: Resultsmentioning

confidence: 99%

Arabidopsis SNARE SYP132 impacts on PIP2;1 trafficking and function in salinity stress

Baena,

Xia,

Waghmare

et al. 2024

The Plant Journal

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SUMMARYIn plants so‐called plasma membrane intrinsic proteins (PIPs) are major water channels governing plant water status. Membrane trafficking contributes to functional regulation of major PIPs and is crucial for abiotic stress resilience. Arabidopsis PIP2;1 is rapidly internalised from the plasma membrane in response to high salinity to regulate osmotic water transport, but knowledge of the underlying mechanisms is fragmentary. Here we show that PIP2;1 occurs in complex with SYNTAXIN OF PLANTS 132 (SYP132) together with the plasma membrane H+‐ATPase AHA1 as evidenced through in vivo and in vitro analysis. SYP132 is a multifaceted vesicle trafficking protein, known to interact with AHA1 and promote endocytosis to impact growth and pathogen defence. Tracking native proteins in immunoblot analysis, we found that salinity stress enhances SYP132 interactions with PIP2;1 and PIP2;2 isoforms to promote redistribution of the water channels away from the plasma membrane. Concurrently, AHA1 binding within the SYP132‐complex was significantly reduced under salinity stress and increased the density of AHA1 proteins at the plasma membrane in leaf tissue. Manipulating SYP132 function in Arabidopsis thaliana enhanced resilience to salinity stress and analysis in heterologous systems suggested that the SNARE influences PIP2;1 osmotic water permeability. We propose therefore that SYP132 coordinates AHA1 and PIP2;1 abundance at the plasma membrane and influences leaf hydraulics to regulate plant responses to abiotic stress signals.

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Section: Resultsmentioning

confidence: 99%