title: ECERIFERUM2-LIKE proteins have unique biochemical and physiological functions in very-long-chain fatty acid elongation.
abstract: The extension of very-long-chain fatty acids (VLCFAs) for the synthesis of specialized apoplastic lipids requires unique biochemical machinery. Condensing enzymes catalyze the first reaction in fatty acid elongation and determine the chain length of fatty acids accepted and produced by the fatty acid elongation complex. Although necessary for the elongation of all VLCFAs, known condensing enzymes cannot efficiently synthesize VLCFAs longer than 28 carbons, despite the prevalence of C28 to C34 acyl lipids in cuticular wax and the pollen coat. The eceriferum2 (cer2) mutant of Arabidopsis (Arabidopsis thaliana) was previously shown to have a specific deficiency in cuticular waxes longer than 28 carbons, and heterologous expression of CER2 in yeast (Saccharomyces cerevisiae) demonstrated that it can modify the acyl chain length produced by a condensing enzyme from 28 to 30 carbon atoms. Here, we report the physiological functions and biochemical specificities of the CER2 homologs CER2-LIKE1 and CER2-LIKE2 by mutant analysis and heterologous expression in yeast. We demonstrate that all three CER2-LIKEs function with the same small subset of condensing enzymes, and that they have different effects on the substrate specificity of the same condensing enzyme. Finally, we show that the changes in acyl chain length caused by each CER2-LIKE protein are of substantial importance for cuticle formation and pollen coat function.
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.).
authors.forename: Tegan M
authors.initials: TM
authors.lastname: Haslam
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.).
authors.forename: Richard
authors.initials: R
authors.lastname: Haslam
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.).
authors.forename: Didier
authors.initials: D
authors.lastname: Thoraval
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.).
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Pascal
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.).
authors.forename: Camille
authors.initials: C
authors.lastname: Delude
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.).
authors.forename: Frédéric
authors.initials: F
authors.lastname: Domergue
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.).
authors.forename: Aurora Mañas
authors.initials: AM
authors.lastname: Fernández
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.).
authors.forename: Frédéric
authors.initials: F
authors.lastname: Beaudoin
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.).
authors.forename: Johnathan A
authors.initials: JA
authors.lastname: Napier
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.) kunst@mail.ubc.ca.
authors.forename: Ljerka
authors.initials: L
authors.lastname: Kunst
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 (T.M.H., A.M.F., L.K.);Department of Biological Chemistry and Crop Protection, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (R.H., F.B., J.A.N.);Université de Bordeaux, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.); andCentre National de la Recherche Scientifique, Laboratoire de Biogenèse Membranaire, Unité Mixte de Recherche 5200, F-33000 Bordeaux, France (D.T., S.P., C.D., F.D., J.J.).
authors.forename: Jérôme
authors.initials: J
authors.lastname: Joubès
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D055432:Metabolomics; D009928:Organ Specificity / Q000235:genetics; D010641:Phenotype; D019441:Plant Epidermis / Q000378:metabolism; D051479:Plant Infertility; D018547:Plant Stems / Q000378:metabolism; D030821:Plants, Genetically Modified; D012098:Reproduction / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013379:Substrate Specificity; D014885:Waxes / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D005227:Fatty Acids; D014885:Waxes