Acylation of non-specific phospholipase C4 determines its function in plant response to phosphate deficiency |
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Authors: | Bao Yang Ke Zhang Xiong Jin Jiayu Yan Shaoping Lu Qingwen Shen Lei Guo Yueyun Hong Xuemin Wang Liang Guo |
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Affiliation: | 1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, 430070 China;2. Department of Biology, University of Missouri, St. Louis, MO, 63121 USA Donald Danforth Plant Science Center, St. Louis, MO, 63132 USA;3. Department of Biology, University of Missouri, St. Louis, MO, 63121 USA |
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Abstract: | Non-specific phospholipase C (NPC) is involved in plant growth, development and stress responses. To elucidate the mechanism by which NPCs mediate cellular functions, here we show that NPC4 is S-acylated at the C terminus and that acylation determines its plasma membrane (PM) association and function. The acylation of NPC4 was detected using NPC4 isolated from Arabidopsis and reconstituted in vitro. The C-terminal Cys-533 was identified as the S-acylation residue, and the mutation of Cys-533 to Ala-533 in NPC4 (NPC4C533A) led to the loss of S-acylation and membrane association of NPC4. The knockout of NPC4 impeded the phosphate deficiency-induced decrease of the phosphosphingolipid glycosyl inositol phosphoryl ceramide (GIPC), but introducing NPC4C533A to npc4-1 failed to complement this defect, thereby supporting the hypothesis that the non-acylated NPC4C533A fails to hydrolyze GIPC during phosphate deprivation. Moreover, NPC4C533A failed to complement the primary root growth in npc4-1 under stress. In addition, NPC4 in Brassica napus was S-acylated and mutation of the S-acylating cysteine residue of BnaC01.NPC4 led to the loss of S-acylation and its membrane association. Together, our results reveal that S-acylation of NPC4 in the C terminus is conserved and required for its membrane association, phosphosphingolipid hydrolysis and function in plant stress responses. |
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Keywords: | Arabidopsis thaliana Brassica napus non-specific phospholipase  C S-acylation membrane association GIPC |
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