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Catalytic activity, duplication and evolution of the CYP98 cytochrome P450 family in wheat

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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.

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Abbreviations

p-CHPLA:

p-coumaroyl-4′-hydroxyphenyllactic acid

P450:

cytochrome P450

THT:

N-(hydroxycinnamoyl)transferase

4-CL:

4-coumaroyl-CoA ligase

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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.

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Authors and Affiliations

  1. Department of Plant Stress Response, Institute of Plant Molecular Biology, CNRS-UPR 2357, Université Louis Pasteur, Centre National de la Recherche Scientifique, 28 rue Goethe, 67000, Strasbourg, France

    Marc Morant, Guillaume A. Schoch, Pascaline Ullmann, Tanya Ertunç, Dawn Little & Danièle Werck-Reichhart

  2. Department of Plant Biology (M.M.) and Department of Chemistry (C.E.O.), The Royal Veterinary and Agricultural University, Thorvaldsensvej 40, DK-1871, Frederiksberg C, Denmark

    Marc Morant & Carl Erik Olsen

  3. Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA, 92037, USA

    Guillaume A. Schoch

  4. Institut fur Pharmazeutische Biologie, Philipps-Universität Marburg, Deutschhausstr. 17A, D-35037, Marburg, Germany

    Maike Petersen

  5. UMR Plante-Microbe-Environnement INRA 1088/CNRS 5184/Université de Bourgogne, BP 86510, 21065 , Dijon Cedex, France

    Jonathan Negrel

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  1. Marc Morant
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  9. Danièle Werck-Reichhart
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Correspondence to Danièle Werck-Reichhart.

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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

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