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Acylcarnitines participate in developmental processes associated to lipid metabolism in plants

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Abstract

Main conclusion

Plant acylcarnitines are present during anabolic processes of lipid metabolism. Their low contents relatively to the corresponding acyl-CoAs suggest that they are associated to specific pools of activated fatty acids.

The non-proteinaceous amino acid carnitine exists in plants either as a free form or esterified to fatty acids. To clarify the biological significance of acylcarnitines in plant lipid metabolism, we have analyzed their content in plant extracts using an optimized tandem mass spectrometry coupled to liquid chromatography method. We have studied different developmental processes (post-germination, organogenesis, embryogenesis) targeted for their high requirement for lipid metabolism. The modulation of the acylcarnitine content was compared to that of the lipid composition and lipid biosynthetic gene expression level in the analyzed materials. Arabidopsis mutants were also studied based on their alteration in de novo fatty acid partitioning between the prokaryotic and eukaryotic pathways of lipid biosynthesis. We show that acylcarnitines cannot specifically be associated to triacylglycerol catabolism but that they are also associated to anabolic pathways of lipid metabolism. They are present during membrane and storage lipid biosynthesis processes. A great divergence in the relative contents of acylcarnitines as compared to the corresponding acyl-CoAs suggests that acylcarnitines are associated to very specific process(es) of lipid metabolism. The nature of their involvement as the transport form of activated fatty acids or in connection with the management of acyl-CoA pools is discussed. Also, the occurrence of medium-chain entities suggests that acylcarnitines are associated with additional lipid processes such as protein acylation for instance. This work strengthens the understanding of the role of acylcarnitines in plant lipid metabolism, probably in the management of specific acyl-CoA pools.

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Abbreviations

ACP:

Acyl carrier protein

CAT:

Carnitine acetyltransferase

CPT:

Carnitine palmitoyltransferase

FA:

Fatty acid

FAT:

Fatty acyl-ACP thioesterase

FFA:

Free fatty acid

KAS:

3-Ketoacyl-ACP synthase

LACS:

Long-chain acyl-CoA synthase

LPAAT:

Lysophosphatidic acid acyltransferase

LPCAT:

Lysophosphatidylcholine acyltransferase

RF:

Response factor

TAG:

Triacylglycerol

WT:

Wild type

References

  1. Abo-Hashema KA, Cake MH, Power GW, Clarke D (1999) Evidence for triacylglycerol synthesis in the lumen of microsomes via a lipolysis-esterification pathway involving carnitine acyltransferases. J Biol Chem 274:35577–35582

  2. Antonenkov VD, Hiltunen JK (2012) Transfer of metabolites across the peroxisomal membrane. Biochim Biophys Acta 1822:1374–1386

  3. Bach L, Gissot L, Marion J, Tellier F, Moreau P, Satiat-Jeunemaître B, Palauqui JC, Napier JA, Faure JD (2011) Very-long-chain fatty acids are required for cell plate formation during cytokinesis in Arabidopsis thaliana. J Cell Sci 124:3223–3234

  4. Bates PD, Stymne S, Ohlrogge J (2013) Biochemical pathways in seed oil synthesis. Curr Opin Plant Biol 16:358–364

  5. Benning C (2009) Mechanisms of lipid transport involved in organelle biogenesis in plant cells. Annu Rev Cell Dev Biol 25:71–91

  6. Bonaventure G, Salas JJ, Pollard MR, Ohlrogge JB (2003) Disruption of the FATB gene in Arabidopsis demonstrates an essential role of saturated fatty acids in plant growth. Plant Cell 15:1020–1033

  7. Bonaventure G, Bao X, Ohlrogge J, Pollard M (2004) Metabolic responses to the reduction in palmitate caused by disruption of the FATB gene in Arabidopsis. Plant Physiol 135:1269–1279

  8. Bourdin B, Adenier H, Perrin Y (2007) Carnitine is associated with fatty acid metabolism in plants. Plant Physiol Biochem 45:926–931

  9. Burgess N, Thomas DR (1986) Carnitine acyltransferase in pea cotyledon mitochondria. Planta 167:58–65

  10. Cruz-Ramírez A, López-Bucio J, Ramírez-Pimentel G, Zurita-Silva A, Sánchez-Calderon L, Ramírez-Chávez E, González-Ortega E, Herrera-Estrella LL (2004) The xipotl mutant of Arabidopsis reveals a critical role for phospholipid metabolism in root system development and epidermal cell integrity. Plant Cell 16:2020–2034

  11. Elgersma Y, van Roermund CW, Wanders RJ, Tabak HF (1995) Peroxisomal and mitochondrial carnitine acetyltransferases of Saccharomyces cerevisiae are encoded by a single gene. EMBO J 14:3472–3479

  12. Ewald R, Hoffmann C, Florian A, Neuhaus E, Fernie AR, Bauwe H (2014) Lipoate-protein ligase and octanoyltransferase are essential for protein lipoylation in mitochondria of Arabidopsis. Plant Physiol 165:978–990

  13. Footitt S, Slocombe SP, Larner V, Kurup S, Wu Y, Larson T, Graham I, Baker A, Holdsworth M (2002) Control of germination and lipid mobilization by COMATOSE, the Arabidopsis homologue of human ALDP. EMBO J 21:2912–2922

  14. Fraenkel G (1953) Studies on the distribution of vitamin BT (carnitine). Biol Bull 104:359–371

  15. Fraser F, Zammit VA (1999) Submitochondrial and subcellular distributions of the carnitine-acylcarnitine carrier. FEBS Lett 445:41–44

  16. Gerbling H, Gerhardt B (1988) Carnitine-acyltransferase activity of mitochondria from mung-bean hypocotyls. Planta 174:90–93

  17. Gooding JM, Shayeghi M, Saggerson ED (2004) Membrane transport of fatty acylcarnitine and free l-carnitine by rat liver microsomes. Eur J Biochem 271:954–961

  18. Graham IA, Eastmond PJ (2002) Pathways of straight and branched chain fatty acid catabolism in higher plants. Prog Lipid Res 41:156–181

  19. Gutierrez L, Mongelard G, Floková K, Pacurar DI, Novák O, Staswick P, Kowalczyk M, Pacurar M, Demailly H, Geiss G, Bellini C (2012) Auxin controls Arabidopsis adventitious root initiation by regulating jasmonic acid homeostasis. Plant Cell 24:2515–2527

  20. Kim S, Yamaoka Y, Ono H, Kim H, Shim D, Maeshima M, Martinoia E, Cahoon EB, Nishida I, Lee Y (2013) AtABCA9 transporter supplies fatty acids for lipid synthesis to the endoplasmic reticulum. Proc Natl Acad Sci USA 110:773–778

  21. Koo AJ, Ohlrogge JB, Pollard M (2004) On the export of fatty acids from the chloroplast. J Biol Chem 279:16101–16110

  22. Kunst L, Browse J, Somerville C (1988) Altered regulation of lipid biosynthesis in a mutant of Arabidopsis deficient in chloroplast glycerol-3-phosphate acyltransferase activity. Proc Natl Acad Sci USA 85:4143–4147

  23. Li N, Gügel IL, Giavalisco P, Zeisler V, Schreiber L, Soll J, Philippar K (2015) FAX1, a novel membrane protein mediating plastid fatty acid export. PLoS Biol 13:e1002053

  24. Li-Beisson Y, Shorrosh B, Beisson F, Andersson MX, Arondel V, Bates PD, Baud S, Bird D, Debono A, Durrett TP, Franke RB, Graham IA, Katayama K, Kelly AA, Larson T, Markham JE, Miquel M, Molina I, Nishida I, Rowland O, Samuels L, Schmid KM, Wada H, Welti R, Xu C, Zallot R, Ohlrogge J (2013) Acyl-lipid metabolism. Arabidopsis Book 11:e0161

  25. Masterson C, Wood C (2000) Pea chloroplast carnitine acetyltransferase. Proc Biol Sci 267:1–6

  26. Masterson C, Wood C (2009) Influence of mitochondrial beta-oxidation on early pea seedling development. New Phytol 181:832–842

  27. McLaren I, Wood C, Jalil MNH, Yong BCS, Thomas DR (1985) Carnitine acyltransferases in chloroplasts of Pisum sativum L. Planta 163:197–200

  28. McNeil PH, Thomas DR (1975) Carnitine content of pea seedling cotyledons. Phytochemistry 14:2335–2336

  29. McNeil PH, Thomas DR (1976) The effect of carnitine on palmitate oxidation by pea cotyledon mitochondria. J Exp Bot 27:1163–1179

  30. Nameth B, Dinka SJ, Chatfield SP, Morris A, English J, Lewis D, Oro R, Raizada MN (2013) The shoot regeneration capacity of excised Arabidopsis cotyledons is established during the initial hours after injury and is modulated by a complex genetic network of light signaling. Plant, Cell Environ 36:68–86

  31. Ohlrogge J, Browse J (1995) Lipid biosynthesis. Plant Cell 7:957–970

  32. Panter RA, Mudd JB (1969) Carnitine levels in some higher plants. FEBS Lett 5:169–170

  33. Ramsay RR, Zammit VA (2004) Carnitine acyltransferases and their influence on CoA pools in health and disease. Mol Aspects Med 25:475–493

  34. Ramsay RR, Gandour RD, Van Der Leij FR (2001) Molecular enzymology of carnitine transfer and transport. Biochem Biophys Acta Protein Struct Mol Enzymol 1546:21–43

  35. Running MP (2014) The role of lipid post-translational modification in plant developmental processes. Front Plant Sci 5:50

  36. Salas JJ, Ohlrogge JB (2002) Characterization of substrate specificity of plant FatA and FatB acyl-ACP thioesterases. Arch Biochem Biophys 403:25–34

  37. Schwabedissen-Gerbling H, Gerhardt B (1995) Purification and characterization of carnitine acyltransferase from higher plant mitochondria. Phytochemistry 39:36–43

  38. Shockey JM, Fulda MS, Browse J (2002) Arabidopsis contains nine long-chain acyl-coenzyme a synthetase genes that participate in fatty acid and glycerolipid metabolism. Plant Physiol 129:1710–1722

  39. Sierra AY, Gratacós E, Carrasco P, Clotet J, Ureña J, Serra D, Asins G, Hegardt FG, Casals N (2008) CPT1c is localized in endoplasmic reticulum of neurons and has carnitine palmitoyltransferase activity. J Biol Chem 283:6878–6885

  40. Steiber A, Kerner J, Hoppel CL (2004) Carnitine: a nutritional, biosynthetic, and functional perspective. Mol Aspects Med 25:455–473

  41. Stephens FB, Constantin-Teodosiu D, Greenhaff PL (2007) New insights concerning the role of carnitine in the regulation of fuel metabolism in skeletal muscle. J Physiol 581:431–444

  42. ter Veld F, Primassin S, Hoffmann L, Mayatepek E, Spiekerkoetter U (2009) Corresponding increase in long-chain acyl-CoA and acylcarnitine after exercise in muscle from VLCAD mice. J Lipid Res 50:1556–1562

  43. Thomas DR, Wood C (1986) The two β-oxidation sites in pea cotyledons carnitine palmitoyltransferase: location and function in pea mitochondria. Planta 168:261–266

  44. Tjellström H, Yang Z, Allen DK, Ohlrogge JB (2012) Rapid kinetic labeling of Arabidopsis cell suspension cultures: implications for models of lipid export from plastids. Plant Physiol 158:601–611

  45. van Roermund CW, Waterham HR, Ijlst L, Wanders RJ (2003) Fatty acid metabolism in Saccharomyces cerevisiae. Cell Mol Life Sci 60:1838–1851

  46. Vrkoslav V, Cvačka J (2012) Identification of the double-bond position in fatty acid methyl esters by liquid chromatography/atmospheric pressure chemical ionisation mass spectrometry. J Chromatogr A 1259:244–250

  47. Wang Z, Benning C (2012) Chloroplast lipid synthesis and lipid trafficking through ER-plastid membrane contact sites. Biochem Soc Trans 40:457–463

  48. Washington L, Cook GA, Mansbach CM 2nd (2003) Inhibition of carnitine palmitoyltransferase in the rat small intestine reduces export of triacylglycerol into the lymph. J Lipid Res 44:1395–1403

  49. Wood C, Jalil MNH, Ariffin A, Yong BCS, Thomas DR (1983) Carnitine short-chain acyltransferases in pea mitochondria. Planta 158:175–178

  50. Wood C, Jalil MNH, McLaren I, Yong BCS, Ariffin A, McNeil PH, Burgess N, Thomas DR (1984) Carnitine long-chain acyltransferase and oxidation of palmitate, palmitoyl CoA and palmitoylcarnitine by pea mitochondria preparations. Planta 161:255–260

  51. Zammit VA, Ramsay RR, Bonomini M, Arduini A (2009) Carnitine, mitochondrial function and therapy. Adv Drug Deliv Rev 61:1353–1362

  52. Zhao L, Katavic V, Li F, Haughn GW, Kunst L (2010) Insertional mutant analysis reveals that long-chain acyl-CoA synthetase 1 (LACS1), but not LACS8, functionally overlaps with LACS9 in Arabidopsis seed oil biosynthesis. Plant J 64:1048–1058

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Acknowledgments

We are grateful to Dr. Laurent Gutierrez and Dr. Stéphanie Guénin from Université de Picardie Jules Verne (Amiens, France) for their assistance in RT-qPCR analyses and Franck Merlier from our group for his technical support in mass spectrometry analysis. We are grateful to Dr. Gustavo Bonaventure from Max Planck Institute (Jena, Germany) for kindly providing with the Arabidopsis mutant genotypes. We also thank a lot Dr. George Lomonossoff from John Innes Centre (Norwich, UK) for taking time to carefully read the manuscript. Finally, we appreciate the time and helpful comments from the reviewers. The work was supported by the French Ministry of National Education, Higher Education and Research.

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Correspondence to Yolande Perrin.

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Nguyen, P., Rippa, S., Rossez, Y. et al. Acylcarnitines participate in developmental processes associated to lipid metabolism in plants. Planta 243, 1011–1022 (2016). https://doi.org/10.1007/s00425-016-2465-y

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Keywords

  • Acyl-CoA
  • Arabidopsis
  • Fatty acid transport
  • Lipid trafficking
  • Plant development
  • Plant metabolism