Unbiased proteomic and forward genetic screens reveal that mechanosensitive ion channel MSL10 functions at ER-plasma membrane contact sites in Arabidopsis thaliana
- Washington University in St. Louis, United States
- SomaLogic, United States
Abstract
Mechanosensitive (MS) ion channels are an evolutionarily conserved way for cells to sense mechanical forces and transduce them into ionic signals. The channel properties of Arabidopsis thaliana MscS-Like (MSL)10 have been well studied, but how MSL10 signals remains largely unknown. To uncover signaling partners of MSL10, we employed both a proteomic screen and a forward genetic screen; both unexpectedly implicated ER-plasma membrane contact sites (EPCSs) in MSL10 function. The proteomic screen revealed that MSL10 associates with multiple proteins associated with EPCSs. Of these, only VAMP-associated proteins (VAP)27-1 and VAP27-3 interacted directly with MSL10. The forward genetic screen, for suppressors of a gain-of-function MSL10 allele (msl10-3G, MSL10S640L), identified mutations in the synaptotagmin (SYT)5 and SYT7 genes. We also found that EPCSs were expanded in leaves of msl10-3G plants compared to the wild type. Taken together, these results indicate that MSL10 associates with and functions with EPCS proteins, providing a new cell-level framework for understanding MSL10 signaling. In addition, placing a mechanosensory protein at EPCSs provides new insight into the function and regulation of this type of subcellular compartment.
Data availability
Mass spectrometry data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD018747, and is included as a Source Data file for Figure 1.
Article and author information
Author details
Richard David Vierstra
Elizabeth S Haswell
Funding
HHMI-Simons Faculty Scholar Grant (55108530)
- Elizabeth S Haswell
National Science Foundation (MCB 1253103)
- Elizabeth S Haswell
National Science Foundation (CMMI-1548571)
- Elizabeth S Haswell
National Science Foundation (DGE-1745038)
- Jennette M Codjoe
William H. Danforth Plant Sciences Fellowship
- Jennette M Codjoe
National Institutes of Health (R01-GM124452)
- Richard David Vierstra
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2022, Codjoe et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Unbiased proteomic and forward genetic screens reveal that mechanosensitive ion channel MSL10 functions at ER-plasma membrane contact sites in Arabidopsis thaliana
eLife 11:e80501.
https://doi.org/10.7554/eLife.80501
