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Functional analyses of differentially expressed isoforms of the Arabidopsis inositol phosphorylceramide synthase

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Abstract

Sphingolipids are key components of eukaryotic plasma membranes that are involved in many functions, including the formation signal transduction complexes. In addition, these lipid species and their catabolites function as secondary signalling molecules in, amongst other processes, apoptosis. The biosynthetic pathway for the formation of sphingolipid is largely conserved. However, unlike mammalian cells, fungi, protozoa and plants synthesize inositol phosphorylceramide (IPC) as their primary phosphosphingolipid. This key step involves the transfer of the phosphorylinositol group from phosphatidylinositol (PI) to phytoceramide, a process catalysed by IPC synthase in plants and fungi. This enzyme activity is at least partly encoded by the AUR1 gene in the fungi, and recently the distantly related functional orthologue of this gene has been identified in the model plant Arabidopsis. Here we functionally analysed all three predicted Arabidopsis IPC synthases, confirming them as aureobasidin A resistant AUR1p orthologues. Expression profiling revealed that the genes encoding these orthologues are differentially expressed in various tissue types isolated from Arabidopsis.

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Abbreviations

AbA:

Aureobasidin A

BSA:

Bovine serum albumin

Cer:

Ceramide

IPC:

Inositol phosphorylceramide

NDB C6-ceramide:

6-((N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino) hexanoyl)sphingosine

PI:

Phosphatidylinositol

SD:

Synthetic minimal media with glucose

SGR:

Synthetic minimal media with galactose

SM:

Sphingomyelin

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Acknowledgments

This work was funded by Biotechnology and Biological Research Council (BB/D52396X/1) and Royal Society (2005/R1) grants to PWD and a British Council/Deutscher Akademischer Austausch Dienst Academic Research Collaboration Award to PWD and RTS. JGM and NKW are funded by the Overseas Research Student Award Scheme. JGM is also funded by the Wolfson Research Institute. This work was also supported in part by the Wolfson Research Institute Collaborative Small Grants Scheme and Deutsche Forschungsgemeinschaft, Bonn.

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Authors and Affiliations

  1. Biophysical Sciences Institute, Department of Chemistry and School of Biological and Biomedical Sciences, Durham University, Durham, UK

    J. G. Mina, N. K. Wansadhipathi-Kannangara, S. Pratt, P. G. Steel & P. W. Denny

  2. School of Medicine and Health, Durham University, Queen’s Campus, Stockton-on-Tees, U.K

    J. G. Mina, N. K. Wansadhipathi-Kannangara, S. Pratt & P. W. Denny

  3. School of Biological and Biomedical Sciences, Durham University, Durham, UK

    Y. Okada & T. Fawcett

  4. Institute for Virology, Medical Center of Hygiene & Medical Microbiology, Philipps-University Marburg, Marburg, Germany

    H. Shams-Eldin & R. T. Schwarz

  5. Unité de Glycobiologie Structurale et Fonctionnelle, Université des Sciences et Technologies de Lille, Lille, France

    R. T. Schwarz

Authors
  1. J. G. Mina
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  2. Y. Okada
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  3. N. K. Wansadhipathi-Kannangara
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  4. S. Pratt
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  5. H. Shams-Eldin
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  6. R. T. Schwarz
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  7. P. G. Steel
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  8. T. Fawcett
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  9. P. W. Denny
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Corresponding authors

Correspondence to T. Fawcett or P. W. Denny.

Additional information

The authors J. G. Mina and Y. Okada contributed equally to this work.

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Mina, J.G., Okada, Y., Wansadhipathi-Kannangara, N.K. et al. Functional analyses of differentially expressed isoforms of the Arabidopsis inositol phosphorylceramide synthase. Plant Mol Biol 73, 399–407 (2010). https://doi.org/10.1007/s11103-010-9626-3

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  • DOI: https://doi.org/10.1007/s11103-010-9626-3

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