Functional characterization of a higher plant sphingolipid Δ4-desaturase

Defining the role of sphingosine and sphingosine-1- phosphate in arabidopsis

Louise V. Michaelson, Simone Zäuner, Jonathan E Markham, Richard P. Haslam, Radhika Desikan, Sarah Mugford, Sandra Albrecht, Dirk Warnecke, Petra Sperling, E. Heinz, Johnathan A. Napier

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

The role of Δ4-unsaturated sphingolipid long-chain bases such as sphingosine was investigated in Arabidopsis (Arabidopsis thaliana). Identification and functional characterization of the sole Arabidopsis ortholog of the sphingolipid Δ4-desaturase was achieved by heterologous expression in Pichia pastoris. A P. pastoris mutant disrupted in the endogenous sphingolipid Δ4-desaturase gene was unable to synthesize glucosylceramides. Synthesis of glucosylceramides was restored by the expression of Arabidopsis gene At4g04930, and these sphingolipids were shown to contain Δ4-unsaturated long-chain bases, confirming that this open reading frame encodes the sphingolipid Δ4-desaturase. At4g04930 has a very restricted expression pattern, transcripts only being detected in pollen and floral tissues. Arabidopsis insertion mutants disrupted in the sphingolipid Δ4-desaturase At4g04930 were isolated and found to be phenotypically normal. Sphingolipidomic profiling of a T-DNA insertion mutant indicated the absence of Δ4-unsaturated sphingolipids in floral tissue, also resulting in the reduced accumulation of glucosylceramides. No difference in the response to drought or water loss was observed between wild-type plants and insertion mutants disrupted in the sphingolipid Δ4-desaturase At4g04930, nor was any difference observed in stomatal closure after treatment with abscisic acid. No differences in pollen viability between wild-type plants and insertion mutants were detected. Based on these observations, it seems unlikely that Δ4-unsaturated sphingolipids and their metabolites such as sphingosine-1-phosphate play a significant role in Arabidopsis growth and development. However, Δ4-unsaturated ceramides may play a previously unrecognized role in the channeling of substrates for the synthesis of glucosylceramides.

Original languageEnglish (US)
Pages (from-to)487-498
Number of pages12
JournalPlant Physiology
Volume149
Issue number1
DOIs
StatePublished - Jan 1 2009

Fingerprint

sphingosine
sphingolipids
Sphingosine
Arabidopsis
Glucosylceramides
Sphingolipids
phosphates
mutants
Pollen
Pichia pastoris
Abscisic Acid
Pichia
Ceramides
Droughts
Growth and Development
pollen
Open Reading Frames
sphingosine 1-phosphate
sphingolipid desaturase
ceramides

ASJC Scopus subject areas

  • Plant Science
  • Genetics
  • Physiology

Cite this

Functional characterization of a higher plant sphingolipid Δ4-desaturase : Defining the role of sphingosine and sphingosine-1- phosphate in arabidopsis. / Michaelson, Louise V.; Zäuner, Simone; Markham, Jonathan E; Haslam, Richard P.; Desikan, Radhika; Mugford, Sarah; Albrecht, Sandra; Warnecke, Dirk; Sperling, Petra; Heinz, E.; Napier, Johnathan A.

In: Plant Physiology, Vol. 149, No. 1, 01.01.2009, p. 487-498.

Research output: Contribution to journalArticle

Michaelson, LV, Zäuner, S, Markham, JE, Haslam, RP, Desikan, R, Mugford, S, Albrecht, S, Warnecke, D, Sperling, P, Heinz, E & Napier, JA 2009, 'Functional characterization of a higher plant sphingolipid Δ4-desaturase: Defining the role of sphingosine and sphingosine-1- phosphate in arabidopsis', Plant Physiology, vol. 149, no. 1, pp. 487-498. https://doi.org/10.1104/pp.108.129411
Michaelson, Louise V. ; Zäuner, Simone ; Markham, Jonathan E ; Haslam, Richard P. ; Desikan, Radhika ; Mugford, Sarah ; Albrecht, Sandra ; Warnecke, Dirk ; Sperling, Petra ; Heinz, E. ; Napier, Johnathan A. / Functional characterization of a higher plant sphingolipid Δ4-desaturase : Defining the role of sphingosine and sphingosine-1- phosphate in arabidopsis. In: Plant Physiology. 2009 ; Vol. 149, No. 1. pp. 487-498.
@article{97635d1ecc614f51baab8555fa0b8b1e,
title = "Functional characterization of a higher plant sphingolipid Δ4-desaturase: Defining the role of sphingosine and sphingosine-1- phosphate in arabidopsis",
abstract = "The role of Δ4-unsaturated sphingolipid long-chain bases such as sphingosine was investigated in Arabidopsis (Arabidopsis thaliana). Identification and functional characterization of the sole Arabidopsis ortholog of the sphingolipid Δ4-desaturase was achieved by heterologous expression in Pichia pastoris. A P. pastoris mutant disrupted in the endogenous sphingolipid Δ4-desaturase gene was unable to synthesize glucosylceramides. Synthesis of glucosylceramides was restored by the expression of Arabidopsis gene At4g04930, and these sphingolipids were shown to contain Δ4-unsaturated long-chain bases, confirming that this open reading frame encodes the sphingolipid Δ4-desaturase. At4g04930 has a very restricted expression pattern, transcripts only being detected in pollen and floral tissues. Arabidopsis insertion mutants disrupted in the sphingolipid Δ4-desaturase At4g04930 were isolated and found to be phenotypically normal. Sphingolipidomic profiling of a T-DNA insertion mutant indicated the absence of Δ4-unsaturated sphingolipids in floral tissue, also resulting in the reduced accumulation of glucosylceramides. No difference in the response to drought or water loss was observed between wild-type plants and insertion mutants disrupted in the sphingolipid Δ4-desaturase At4g04930, nor was any difference observed in stomatal closure after treatment with abscisic acid. No differences in pollen viability between wild-type plants and insertion mutants were detected. Based on these observations, it seems unlikely that Δ4-unsaturated sphingolipids and their metabolites such as sphingosine-1-phosphate play a significant role in Arabidopsis growth and development. However, Δ4-unsaturated ceramides may play a previously unrecognized role in the channeling of substrates for the synthesis of glucosylceramides.",
author = "Michaelson, {Louise V.} and Simone Z{\"a}uner and Markham, {Jonathan E} and Haslam, {Richard P.} and Radhika Desikan and Sarah Mugford and Sandra Albrecht and Dirk Warnecke and Petra Sperling and E. Heinz and Napier, {Johnathan A.}",
year = "2009",
month = "1",
day = "1",
doi = "10.1104/pp.108.129411",
language = "English (US)",
volume = "149",
pages = "487--498",
journal = "Plant Physiology",
issn = "0032-0889",
publisher = "American Society of Plant Biologists",
number = "1",

}

TY - JOUR

T1 - Functional characterization of a higher plant sphingolipid Δ4-desaturase

T2 - Defining the role of sphingosine and sphingosine-1- phosphate in arabidopsis

AU - Michaelson, Louise V.

AU - Zäuner, Simone

AU - Markham, Jonathan E

AU - Haslam, Richard P.

AU - Desikan, Radhika

AU - Mugford, Sarah

AU - Albrecht, Sandra

AU - Warnecke, Dirk

AU - Sperling, Petra

AU - Heinz, E.

AU - Napier, Johnathan A.

PY - 2009/1/1

Y1 - 2009/1/1

N2 - The role of Δ4-unsaturated sphingolipid long-chain bases such as sphingosine was investigated in Arabidopsis (Arabidopsis thaliana). Identification and functional characterization of the sole Arabidopsis ortholog of the sphingolipid Δ4-desaturase was achieved by heterologous expression in Pichia pastoris. A P. pastoris mutant disrupted in the endogenous sphingolipid Δ4-desaturase gene was unable to synthesize glucosylceramides. Synthesis of glucosylceramides was restored by the expression of Arabidopsis gene At4g04930, and these sphingolipids were shown to contain Δ4-unsaturated long-chain bases, confirming that this open reading frame encodes the sphingolipid Δ4-desaturase. At4g04930 has a very restricted expression pattern, transcripts only being detected in pollen and floral tissues. Arabidopsis insertion mutants disrupted in the sphingolipid Δ4-desaturase At4g04930 were isolated and found to be phenotypically normal. Sphingolipidomic profiling of a T-DNA insertion mutant indicated the absence of Δ4-unsaturated sphingolipids in floral tissue, also resulting in the reduced accumulation of glucosylceramides. No difference in the response to drought or water loss was observed between wild-type plants and insertion mutants disrupted in the sphingolipid Δ4-desaturase At4g04930, nor was any difference observed in stomatal closure after treatment with abscisic acid. No differences in pollen viability between wild-type plants and insertion mutants were detected. Based on these observations, it seems unlikely that Δ4-unsaturated sphingolipids and their metabolites such as sphingosine-1-phosphate play a significant role in Arabidopsis growth and development. However, Δ4-unsaturated ceramides may play a previously unrecognized role in the channeling of substrates for the synthesis of glucosylceramides.

AB - The role of Δ4-unsaturated sphingolipid long-chain bases such as sphingosine was investigated in Arabidopsis (Arabidopsis thaliana). Identification and functional characterization of the sole Arabidopsis ortholog of the sphingolipid Δ4-desaturase was achieved by heterologous expression in Pichia pastoris. A P. pastoris mutant disrupted in the endogenous sphingolipid Δ4-desaturase gene was unable to synthesize glucosylceramides. Synthesis of glucosylceramides was restored by the expression of Arabidopsis gene At4g04930, and these sphingolipids were shown to contain Δ4-unsaturated long-chain bases, confirming that this open reading frame encodes the sphingolipid Δ4-desaturase. At4g04930 has a very restricted expression pattern, transcripts only being detected in pollen and floral tissues. Arabidopsis insertion mutants disrupted in the sphingolipid Δ4-desaturase At4g04930 were isolated and found to be phenotypically normal. Sphingolipidomic profiling of a T-DNA insertion mutant indicated the absence of Δ4-unsaturated sphingolipids in floral tissue, also resulting in the reduced accumulation of glucosylceramides. No difference in the response to drought or water loss was observed between wild-type plants and insertion mutants disrupted in the sphingolipid Δ4-desaturase At4g04930, nor was any difference observed in stomatal closure after treatment with abscisic acid. No differences in pollen viability between wild-type plants and insertion mutants were detected. Based on these observations, it seems unlikely that Δ4-unsaturated sphingolipids and their metabolites such as sphingosine-1-phosphate play a significant role in Arabidopsis growth and development. However, Δ4-unsaturated ceramides may play a previously unrecognized role in the channeling of substrates for the synthesis of glucosylceramides.

UR - http://www.scopus.com/inward/record.url?scp=58449095434&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=58449095434&partnerID=8YFLogxK

U2 - 10.1104/pp.108.129411

DO - 10.1104/pp.108.129411

M3 - Article

VL - 149

SP - 487

EP - 498

JO - Plant Physiology

JF - Plant Physiology

SN - 0032-0889

IS - 1

ER -