Acylation of non‐specific phospholipase C4 determines its function in plant response to phosphate deficiency

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 (NPC4 C533A ) 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 NPC4 C533A to npc4-1 failed to complement this defect, thereby supporting the hypothesis that the non-acylated NPC4 C533A fails to hydrolyze GIPC during phosphate deprivation. Moreover, NPC4 C533A 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.

Validating a Major Quantitative Trait Locus and Predicting Candidate Genes Associated With Kernel Width Through QTL Mapping and RNA-Sequencing Technology Using Near-Isogenic Lines in Maize

“…After DEG analysis, a total of 12 protein-coding genes were differentially expressed and retained in the qKW-1 physical intervals ( Table 6 ). Previous studies indicated that non-specific phospholipase C (NPC) is involved in plant growth, development, and stress responses ( Yang et al, 2021 ). NRT1.1B is associated with root microbiota composition and nitrogen use in field-grown rice ( Zhang et al, 2019 ).…”

“…Finally, 12 DEGs were obtained, and the 12 genes were used for comparative genomics studies. Previous studies indicated that non-specific phospholipase C is involved in plant growth, development and stress responses ( Yang et al, 2021 ). Results from Cao et al (2016) indicate that the overexpression of NPC1 leads to brittle stem nodes and panicle nodes and increased seed shattering due to a decrease in mechanical strength in nodes tissues.…”

Pyruvate transporter BnaBASS2 impacts seed oil accumulation in Brassica napus

“…The CDS of BnaA05.BASS2 was amplified and ligated into the pMDC83 vector, which contains a 2 × 35S promoter and a C‐terminal GFP tag. The recombinant BnaA05.BASS2::GFP vector was transformed into the mesophyll protoplasts of Arabidopsis (Yoo et al ., 2007 ) and epidermal cells of tobacco (Yang et al ., 2021 ). The fluorescence pictures were observed using a Leica confocal microscope at 12 h after transfection.…”

Screening DHHCs of S‐acylated proteins using an OsDHHCcDNA library and bimolecular fluorescence complementation in rice

“…The receptor‐like cytoplasmic kinases (RLCKs) Lost In Pollen tube guidance 1 (LIP1) and LIP2 are double S ‐acylated and help guide pollen tube growth to the ovule (Liu et al., 2013). S ‐acylation of non‐specific phospholipase C4 (NPC4) is required for its membrane association, phosphosphingolipid hydrolysis, and function in the plant stress response (Yang et al., 2021).…”