Arabidopsis 56–Amino Acid Serine Palmitoyltransferase- Interacting Proteins Stimulate Sphingolipid Synthesis, Are Essential, and Affect Mycotoxin Sensitivity

Authors

Athen N. Kimberlin, University of Nebraska-LincolnFollow
Saurav Majumder, Uniformed Services University of the Health Sciences
Gongshe Han, Uniformed Services University of the Health Sciences
Ming Chen, University of Nebraska-Lincoln
Rebecca E. Cahoon, University of Nebraska-LincolnFollow
Julie M. Stone, University of Nebraska-LincolnFollow
Teresa M. Dunn, Uniformed Services University of the Health Sciences
Edgar B. Cahoon, University of Nebraska-LincolnFollow

Document Type

Article

Abstract

Maintenance of sphingolipid homeostasis is critical for cell growth and programmed cell death (PCD). Serine palmitoyltransferase (SPT), composed of LCB1 and LCB2 subunits, catalyzes the primary regulatory point for sphingolipid synthesis. Small subunits of SPT (ssSPT) that strongly stimulate SPT activity have been identified in mammals, but the role of ssSPT in eukaryotic cells is unclear. Candidate Arabidopsis thaliana ssSPTs, ssSPTa and ssSPTb, were identified and characterized. Expression of these 56–amino acid polypeptides in a Saccharomyces cerevisiae SPT null mutant stimulated SPT activity from the Arabidopsis LCB1/LCB2 heterodimer by >100-fold through physical interaction with LCB1/LCB2. ssSPTa transcripts were more enriched in all organs and >400-fold more abundant in pollen than ssSPTb transcripts. Accordingly, homozygous ssSPTa T-DNA mutants were not recoverable, and 50% nonviable pollen was detected in heterozygous ssspta mutants. Pollen viability was recovered by expression of wild-type ssSPTa or ssSPTb under control of the ssSPTa promoter, indicating ssSPTa and ssSPTb functional redundancy. SPT activity and sensitivity to the PCD-inducing mycotoxin fumonisin B₁ (FB₁) were increased by ssSPTa overexpression. Conversely, SPT activity and FB₁ sensitivity were reduced in ssSPTa RNA interference lines. These results demonstrate that ssSPTs are essential for male gametophytes, are important for FB₁ sensitivity, and limit sphingolipid synthesis in planta.