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Molecular and biochemical characterization of two brassinosteroid sulfotransferases from Arabidopsis, AtST4a (At2g14920) and AtST1 (At2g03760)

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Abstract

Mammalian sulfotransferases (EC 2.8.2) are involved in many important facets of steroid hormone activity and metabolism. In this study, Arabidopsis AtST4a and AtST1 were identified and characterized as brassinosteroid sulfotransferases that appear to be involved in different aspects of hormone regulation. The two proteins share 44% identity in amino acid sequence, and belong to different plant sulfotransferase families. AtST4a was specific for biologically active end products of the brassinosteroid pathway. The enzyme sulfated brassinosteroids with diverse side-chain structures, including 24-epibrassinosteroids and the naturally occurring (22R, 23R)-28-homobrassinosteroids. AtST4a belongs to a small subfamily of sulfotransferases having two other members, AtST4b and -c. Among the three recombinant enzymes, only AtST4a was catalytically active with brassinosteroids. Transcript expression of AtST4 subfamily members was largely specific to the root. AtST4b- and -c transcript levels were induced by treatment with trans-zeatin, while AtST4a was repressed under the same conditions, supporting a divergent function of AtST4a. Co-regulation of AtST4b and -c correlated with their location in tandem on chromosome 1. AtST1 was stereospecific for 24-epibrassinosteroids, with a substrate preference for the metabolic precursor 24-epicathasterone, and exhibited catalytic activity with hydroxysteroids and estrogens. To gain more insight into this dual activity with plant and mammalian steroids, enzymatic activities of human steroid sulfotransferases toward brassinosteroids were characterized. The dehydroepiandrosterone sulfotransferase SULT2A1 displayed catalytic activity with a selected set of 24-epibrassinolide precursors, including 24-epicathasterone, with specific activities comparable to that measured for the endogenous substrate dehydroepiandrosterone. The comparable activity profiles of AtST1 and SULT2A1 suggest a similar architecture of the acceptor-binding site between the two enzymes, and may potentially reflect a common ability to conjugate certain xenobiotics.

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Abbreviations

TAIR:

The arabidopsis information resource

IPTG:

Isopropyl β-d-thiogalactopyranoside

PAPS:

3′-Phosphoadenosine 5′-phosphosulfate

DON:

Deoxynivalenol

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Acknowledgments

We thank Dr. G. Adam (Leibniz Institute of Plant Biochemistry, Halle, Germany) for brassinosteroid substrates, Dr. D. Roby (Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique, Castanet-Tolosan, France) for the gift of the AtST1 cDNA (RaR047), Dr. C. N. Falany (University of Alabama at Birmingham, AL, USA) for the gift of the human SULT1E1 cDNA, Dr. V. Luu-The (Centre de recherche du CHUL, Sainte-Foy, QU, Canada) for the gift of the human SULT2A1 cDNA, and Drs. Yuzuki Manabe and Brian Miki (Agriculture and Agri-Food Canada, Eastern Cereal and Oilseed Research Centre, Ottawa, ON, Canada) for sharing data prior to publication. We also thank Alex Molnar for assistance in preparing the figures. F. M. was supported in part by a postgraduate scholarship from the Natural Sciences and Engineering Research Council (NSERC) of Canada, and a Concordia J. W. McConnell Memorial graduate fellowship. This work was supported by NSERC.

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  1. Agriculture and Agri-Food Canada, Southern Crop Protection and Food Research Centre, 1391 Sandford St., London, ON, N5V 4T3, Canada

    Frédéric Marsolais

  2. Centre for Structural and Functional Genomics, Department of Biology, Concordia University, 7141 Sherbrooke W., H4A 1R6, Montréal, QC, Canada

    Frédéric Marsolais, Jason Boyd, Yosabeth Paredes, Anna-Maria Schinas, Melina Garcia, Samar Elzein & Luc Varin

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Correspondence to Frédéric Marsolais.

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Marsolais, F., Boyd, J., Paredes, Y. et al. Molecular and biochemical characterization of two brassinosteroid sulfotransferases from Arabidopsis, AtST4a (At2g14920) and AtST1 (At2g03760). Planta 225, 1233–1244 (2007). https://doi.org/10.1007/s00425-006-0413-y

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