Sphingolipid Long-Chain Base Phosphate Degradation Can Be a Rate-Limiting Step in Long-Chain Base Homeostasis

Benjamin Lambour; René Glenz; Carmen Forner; Markus Krischke; Martin J Mueller; Agnes Fekete; Frank Waller

doi:10.3389/fpls.2022.911073

Sphingolipid Long-Chain Base Phosphate Degradation Can Be a Rate-Limiting Step in Long-Chain Base Homeostasis

Front Plant Sci. 2022 Jun 15:13:911073. doi: 10.3389/fpls.2022.911073. eCollection 2022.

Authors

Benjamin Lambour¹, René Glenz¹, Carmen Forner¹, Markus Krischke¹, Martin J Mueller¹, Agnes Fekete¹, Frank Waller¹

Affiliation

¹ Pharmaceutical Biology, Julius-von-Sachs Institute of Biosciences, Julius-Maximilians-Universität Würzburg, Würzburg, Germany.

Abstract

Sphingolipid long-chain bases (LCBs) are building blocks for membrane-localized sphingolipids, and are involved in signal transduction pathways in plants. Elevated LCB levels are associated with the induction of programmed cell death and pathogen-derived toxin-induced cell death. Therefore, levels of free LCBs can determine survival of plant cells. To elucidate the contribution of metabolic pathways regulating high LCB levels, we applied the deuterium-labeled LCB D-erythro-sphinganine-d7 (D₇-d18:0), the first LCB in sphingolipid biosynthesis, to Arabidopsis leaves and quantified labeled LCBs, LCB phosphates (LCB-Ps), and 14 abundant ceramide (Cer) species over time. We show that LCB D₇-d18:0 is rapidly converted into the LCBs d18:0P, t18:0, and t18:0P. Deuterium-labeled ceramides were less abundant, but increased over time, with the highest levels detected for Cer(d18:0/16:0), Cer(d18:0/24:0), Cer(t18:0/16:0), and Cer(t18:0/22:0). A more than 50-fold increase of LCB-P levels after leaf incubation in LCB D₇-d18:0 indicated that degradation of LCBs via LCB-Ps is important, and we hypothesized that LCB-P degradation could be a rate-limiting step to reduce high levels of LCBs. To functionally test this hypothesis, we constructed a transgenic line with dihydrosphingosine-1-phosphate lyase 1 (DPL1) under control of an inducible promotor. Higher expression of DPL1 significantly reduced elevated LCB-P and LCB levels induced by Fumonisin B₁, and rendered plants more resistant against this fungal toxin. Taken together, we provide quantitative data on the contribution of major enzymatic pathways to reduce high LCB levels, which can trigger cell death. Specifically, we provide functional evidence that DPL1 can be a rate-limiting step in regulating high LCB levels.

Keywords: LC–MS/MS; cell death; dihydrosphingosine-1-phosphate lyase; long-chain base; metabolic flux analysis; plant sphingolipid metabolism; sphingolipid.