Abstract
A burst of evolutionary duplication upon land colonization seems to have led to the large superfamily of cytochromes P450 in higher plants. Within this superfamily some clans and families are heavily duplicated. Others, such as genes involved in the phenylpropanoid pathway have led to fewer duplication events. Eight coding sequences belonging to the CYP98 family reported to catalyze the 3-hydroxylation step in this pathway were isolated from Triticum aestivum (wheat) and expressed in yeast. Comparison of the catalytic properties of the recombinant enzymes with those of CYP98s from other plant taxa was coupled to phylogenetic analyses. Our results indicate that the unusually high frequency of gene duplication in the wheat CYP98 family is a direct or indirect result from ploidization. While ancient duplication led to evolution of enzymes with different substrate preferences, most of recent duplicates underwent silencing via degenerative mutations. Three of the eight tested CYP98s from wheat have phenol meta-hydroxylase activity, with p-coumaroylshikimate being the primary substrate for all of these, as it is the case for CYP98s from sweet basil and Arabidopsis thaliana. However, CYP98s from divergent taxa have acquired different additional subsidiary activities. Some of them might be significant in the metabolism of various free or conjugated phenolics in different plant species. One of the most significant is meta-hydroxylation of p-coumaroyltyramine, predominantly by the wheat enzymes, for the synthesis of suberin phenolic monomers. Homology modeling, confirmed by directed mutagenesis, provides information on the protein regions and structural features important for some observed changes in substrate selectivity. They indicate that the metabolism of quinate ester and tyramine amide of p-coumaric acid rely on the same recognition site in the protein.
Similar content being viewed by others
Abbreviations
- p-CHPLA:
-
p-coumaroyl-4′-hydroxyphenyllactic acid
- P450:
-
cytochrome P450
- THT:
-
N-(hydroxycinnamoyl)transferase
- 4-CL:
-
4-coumaroyl-CoA ligase
References
Abdulrazzak N, Pollet B, Ehlting J, Larsen K, Asnaghi C, Ronseau S, Proux C, Erhardt M, Seltzer V, Renou JP, Ullmann P, Pauly M, Lapierre C, Werck-Reichhart D (2006) A coumaroyl-ester-3-hydroxylase insertion mutant reveals the existence of nonredundant meta-hydroxylation pathways and essential roles for phenolic precursors in cell expansion and plant growth. Plant Physiol 140:30–48
Berman HM, Westbrook J, Feng Z, Gilliland G, Bhat TN, Weissig H, Shindyalov IN, Bourne PE (2000) The Protein Data Bank. Nucleic Acids Res 28:235–242
Bernards MA (2002) Demystifying suberin. Can J Bot 80:227–240
Bouchereau A, Aziz A, Larher F, Martin-Tanguy J (1999) Polyamines and environmental challenges: recent development. Plant Sci 140:103–125
Brugliera F, Barri-Rewell G, Holton TA, Mason JG (1999) Isolation and characterization of a flavonoid 3’-hydroxylase cDNA clone corresponding to the Ht1 locus of Petunia hybrida. Plant J 19:441–451
Brunger AT, Adams PD, Clore GM, DeLano WL, Gros P, Grosse-Kunstleve R W, Jiang J.S, Kuszewski J, Nilges M, Pannu NS, Read RJ, Rice LM, Simonson T, Warren GL (1998). Crystallography & NMR system: a new software suite for macromolecular structure determination. Acta Crystallogr D Biol Crystallogr 54:905–921
Bundock PC, Christopher JT, Eggler P, Ablett G, Henry RJ, Holton TA (2003). Single nucleotide polymorphisms in cytochrome P450 genes from barley. Theor Appl Genet 106:676–682
Cabello-Hurtado F, Zimmerlin A, Rahier A, Taton M, DeRose R, Nedelkina S, Batard Y, Durst F, Pallet KE, Werck-Reichhart D (1997) Cloning and functional expression in yeast of a cDNA coding for an obtusifoliol 14α-demethylase (CYP51) in wheat. Biochem Biophys Res Commun 230:381–385
Devos KM, Gale MD (2000) Genome relationships: the grass model in current research. Plant Cell 12:637–646
Dickmann LJ, Locuson CW, Jones JP, Rettie AE (2004) The roles of Arg97, Asp293, and Arg108 in enzyme stability and substrate specificity of CYP2C9. Mol Pharmacol 65:842–850
Eckardt NA (2001) A sense of self: the role of DNA allopolyploidization. Plant Cell 13:1699–1704
Ehlting J, Buttner D, Wang Q, Douglas CJ, Somssich IE, Kombrink E (1999) Three 4-coumarate:coenzyme A ligases in Arabidopsis thaliana represent two evolutionarily divergent classes in angiosperms. Plant J 19:9–20
Ertunç T (2002) Metabolismus des Fungizids Cyprodinil in Weizen sowie Aufklärung der Struktur- und Bindungsformen nicht-extrahierbarer Rückstände mittels spektroskopischer und chromatographischer Methoden; Verlag Mainz, Aachen, Germany. ISBN 3-86130-068-0
Facchini PJ, Hagel J, Zulak KG (2002) Hydroxycinnamic acid amide metabolism: physiology and biochemistry. Can J Bot 80:577–589
Franke R, Hemm MR, Denault JW, Ruegger MO, Humphreys JM, Chapple C (2002a) Changes in secondary metabolism and deposition of an unusual lignin in the ref8 mutant of Arabidopsis. Plant J 30:47–59
Franke R, Humphreys JM, Hemm MR, Denault JW, Ruegger MO, Cusumano JC, Chapple C (2002b) The Arabidopsis REF8 gene encodes the 3-hydroxylase of phenylpropanoid metabolism. Plant J 30:33–45
Frear S (1995) Wheat microsomal cytochrome P450 monooxygenases: characterization and importance in the metabolic detoxification and selectivity of wheat herbicides. Drug Metab Drug Interact 12:329–358
Gang DR, Beuerle T, Ullmann P, Werck-Reichhart D, Pichersky E (2002) Differential production of meta hydroxylated phenylpropanoids in sweet basil peltate glandular trichomes and leaves is controlled by the activities of specific acyltransferases and hydroxylases. Plant Physiol 130:1536–1544
Gotoh O (1992) Substrate recognition sites in cytochrome P450 family 2 (CYP2) proteins inferred from comparative analyses of amino acid and coding nucleotide sequences. J Biol Chem 267:83–90
Grace SC, Logan BA (2000) Energy dissipation and radical scavenging by the plant phenylpropanoid pathway. Philos Trans R Soc Lond B Biol Sci 355:1499–1510
Guillet G, De Luca V (2005) Wound-inducible biosynthesis of phytoalexin hydroxycinnamic acid amides of tyramine in tryptophan and tyrosine decarboxylase transgenic tobacco lines. Plant Physiol 137:692–699
Hagel JM, Facchini PJ (2005) Elevated tyrosine decarboxylase and tyramine hydroxycinnamoyltransferase levels increase wound-induced tyramine-derived hydroxycinnamic acid amide accumulation in transgenic tobacco leaves. Planta 221:904–914
Hehn A, Morant M, Werck-Reichhart D (2002) Partial recoding of P450 and P450 reductase cDNAs for improved expression in yeast and plants. Methods Enzymol 357:343–351
Heller W, Kühnl T (1985) Elicitor induction of a microsomal 5-O-(4-coumaroyl)shikimate 3’-hydroxylase in parsley cell suspension cultures. Arch Biochem Biophys 241:453–460
Hoffmann L, Maury S, Martz F, Geoffroy P, Legrand M (2003) Purification, cloning, and properties of an acyltransferase controlling shikimate and quinate ester intermediates in phenylpropanoid metabolism. J Biol Chem 278:95–103
Holton TA, Brugliera F, Lester DR, Tanaka Y, Hyland CD, Menting JGT, Lu CY, Farcy E, Stevenson TW, Cornish EC (1993) Cloning and expression of cytochrome P450 genes controlling flower colour. Nature 366:276–279
Ishihara A, Kawata N, Matsukawa T, Iwamura H (2000) Induction of N-hydroxycinnamoyltyramine synthesis and tyramine N-hydroxycinnamoyltransferase (THT) activity by wounding in maize leaves. Biosci Biotechnol Biochem 64:1025–1031
Jones TA, Kjeldgaard M (1997) Electro-density map interpretation. Methods Enzymol 277:173–208
Jørgensen K, Rasmussen AV, Morant M, Nielsen AH, Bjarnholt N, Zagrobelny M, Bak S, Møller BL (2005) Metabolon formation and metabolic channeling in the biosynthesis of plant natural products. Curr Op Plant Biol 8:280–291
Kühnl T, Koch U, Heller W, Wellmann E (1987) Chlorogenic acid biosynthesis: characterization of a light-induced microsomal 5-O-(4-coumaroyl)-D-quinate/shikimate 3’-hydroxylase from carrot (Daucus carota L.) cell suspension cultures. Arch Biochem Biophys 258:226–232
Lynch M, Conery JS (2000) The evolutionary fate and consequences of duplicate genes. Science 290:1151–1155
Lynch M, Force A (2000) The probability of duplicate gene preservation by subfunctionalization. Genetics 154:459–473
Matsuno M, Nagatsu A, Ogihara Y, Ellis BE, Mizukami H (2002) CYP98A6 from Lithospermum erythrorhizon encodes 4-coumaroyl-4’-hydroxyphenyllactic acid 3-hydroxylase involved in rosmarinic acid biosynthesis. FEBS Lett 514:219–224
Morant M, Hehn A, Werck-Reichhart D (2002) Conservation and diversity of gene families explored using the CODEHOP strategy in higher plants. BMC Plant Biol 2:7
Nair RB, Xia Q, Kartha CJ, Kurylo E, Hirji RN, Datla R, Selvaraj G (2002) Arabidopsis CYP98A3 mediating aromatic 3-hydroxylation. developmental regulation of the gene, and expression in yeast. Plant Physiol 130:210–220
Negrel J, Javelle F (1997) Purification, characterization and partial amino acid sequencing of hydroxycinnamoyl-CoA:tyramine N-(hydroxycinnamoyl)transferase from tobacco cell-suspension cultures. Eur J Biochem 247:1127–1135
Newman MA, von Roepenack-Lahaye E, Parr A, Daniels MJ, Dow JM (2001) Induction of hydroxycinnamoyl-tyramine conjugates in pepper by Xanthomonas campestris, a plant defense response activated by hrp gene-dependent and hrp gene-independent mechanisms. Mol Plant Microbe Interact 14:785–792
Nomura T, Ishihara A, Imaishi H, Ohkawa H, Endo TR, Iwamura H (2003) Rearrangement of the genes for the biosynthesis of benzoxazinones in the evolution of Triticeae species. Planta 217:776–782
Omura T, Sato R (1964) The carbon monoxide binding pigment of liver microsomes. I. Evidence for its hemoprotein nature. J Biol Chem 239:2370–2378
Ozkan H, Levy AA, Feldman M (2001) Allopolyploidy-induced rapid genome evolution in the wheat (Aegilops-Triticum) group. Plant Cell 13:1735–1747
Paquette SM, Bak S, Feyereisen R (2000) Intron-exon organization and phylogeny in a large superfamily, the paralogous cytochrome P450 genes of Arabidopsis thaliana. DNA Cell Biol 19:307–317
Petersen M (1997) Cytochrome P-450-dependent hydroxylation in the biosynthesis of rosmarinic acid in Coleus. Phytochemistry 45:1165–1172
Petersen M, Simmonds MS (2003) Rosmarinic acid. Phytochemistry 62:121–125
Pompon D, Louerat B, Bronine A, Urban P (1996) Yeast expression of animal and plant P450s in optimized redox environments. Methods Enzymol 272:51–64
Sapp M, Ertunç T, Bringmann I, Schäffer A, Schmidt B (2004) Characterization of the bound residues of the fungicide cyprodinil formed in plant cell suspension cultures of wheat. Pest Manage Sci 60(1):65–74
Schmidt A, Grimm R, Schmidt J, Scheel D, Strack D, Rosahl S (1999) Cloning and expression of a potato cDNA encoding hydroxycinnamoyl-CoA:tyramine N-(hydroxycinnamoyl) transferase. J Biol Chem 274:4273–4280
Schoch G, Goepfert S, Morant M, Hehn A, Meyer D, Ullmann P, Werck-Reichhart D (2001) CYP98A3 from Arabidopsis thaliana is a 3’-hydroxylase of phenolic esters, a missing link in the phenylpropanoid pathway. J Biol Chem 276:36566–36574
Schoch GA, Yano JK, Wester MR, Griffin KJ, Stout CD, Johnson EF (2004) Structure of human microsomal cytochrome P450 2C8. Evidence for a peripheral fatty acid binding site. J Biol Chem 279:9497–9503
Schoenbohm C, Martens S, Eder C, Forkmann G, Weisshaar B (2000) Identification of the Arabidopsis thaliana flavonoid 3’-hydroxylase gene and functional expression of the encoded P450 enzyme. Biol Chem 381:749–753
Schuettelkopf W, van Aalten DMF (2004) PRODRG – a tool for high-throughput crystallography of protein-ligand complexes. Acta Crystallogr D 60:1355–1363
Schwede T, Kopp J, Guex N, Peitsch MC (2003) SWISS-MODEL: an automated protein homology-modeling server. Nucleic Acids Res 31:3381–3385
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882
Villegas M, Brodelius PE (1990) Elicitor-induced hydroxycinnamoyl-CoA: tyramine hydroxycinnamoyltransferase in plant cell suspension cultures. Physiol Plant 78:414–420
Wester MR, Yano JK, Schoch GA, Yang C, Griffin KJ, Stout CD, Johnson EF (2004) The structure of human cytochrome P450 2C9 complexed with flurbiprofen at 2.0-Å resolution. J Biol Chem 279:35630–35637
Word JM, Lovell SC, Richardson JS, Richardson DC (1999) Asparagine and glutamine: using hydrogen atom contacts in the choice of side-chain amide orientation. J Mol Biol 285:1735–1747
Xu F, Lagudah ES, Moose SP, Riechers DE (2002) Tandemly duplicated safener-induced glutathione S-transferase genes from Triticum tauschii contribute to genome- and organ-specific expression in hexaploid wheat. Plant Physiol 130:362–373
Acknowledgements
We thank Pr. David Nelson (University of Tennessee, Memphis) and Dr. Suzanne Paquette (University of Washington, Seattle) for their advices and contribution to the naming of the new P450s and the phylogenetic analysis. The WAT11 yeast strain and pYeDP60 vector were provided by Dr. Denis Pompon (Gif-sur-Yvette, France), the CYP98A13v1 and CYP98A13v2 cDNAs by Dr. David Gang (University of Arizona, Tucson). Technical help of A. Lesot was appreciated. Support of Bayer CropScience and of the Association Nationale de la Recherche Technique to M.M. are gratefully acknowledged. We are very grateful to Jürgen Ehlting for his critical reading of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Morant, M., Schoch, G.A., Ullmann, P. et al. Catalytic activity, duplication and evolution of the CYP98 cytochrome P450 family in wheat . Plant Mol Biol 63, 1–19 (2007). https://doi.org/10.1007/s11103-006-9028-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11103-006-9028-8