Compared to animals, evolution of plant calcium (Ca) physiology has led to a loss of proteins for influx and small ligand-operated control of cytosolic Ca, leaving many Ca mechanisms unaccounted for. Here, we show a mechanism for sorting and activation of glutamate receptor-like channels (GLRs) by CORNICHON HOMOLOG (CNIH) proteins. Single mutants of pollen-expressed GLRs (GLRs) showed growth and Ca flux phenotypes expected for plasma membrane Ca channels. However, higher-order mutants of GLR3.3 revealed phenotypes contradicting this assumption. These discrepancies could be explained by subcellularGLR localization, and we explored the implication of CNIHs in this sorting. We found thatGLRs interact with CNIH pairs, yielding specific intracellular localizations.CNIHs further trigger GLR activity in mammalian cells without any ligand. These results reveal a regulatory mechanism underlying Ca homeostasis by sorting and activation of GLRs byCNIHs.
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