title: Chemical Genetics Uncovers Novel Inhibitors of Lignification, Including p-Iodobenzoic Acid Targeting CINNAMATE-4-HYDROXYLASE.
abstract: Plant secondary-thickened cell walls are characterized by the presence of lignin, a recalcitrant and hydrophobic polymer that provides mechanical strength and ensures long-distance water transport. Exactly the recalcitrance and hydrophobicity of lignin put a burden on the industrial processing efficiency of lignocellulosic biomass. Both forward and reverse genetic strategies have been used intensively to unravel the molecular mechanism of lignin deposition. As an alternative strategy, we introduce here a forward chemical genetic approach to find candidate inhibitors of lignification. A high-throughput assay to assess lignification in Arabidopsis (Arabidopsis thaliana) seedlings was developed and used to screen a 10-k library of structurally diverse, synthetic molecules. Of the 73 compounds that reduced lignin deposition, 39 that had a major impact were retained and classified into five clusters based on the shift they induced in the phenolic profile of Arabidopsis seedlings. One representative compound of each cluster was selected for further lignin-specific assays, leading to the identification of an aromatic compound that is processed in the plant into two fragments, both having inhibitory activity against lignification. One fragment, p-iodobenzoic acid, was further characterized as a new inhibitor of CINNAMATE 4-HYDROXYLASE, a key enzyme of the phenylpropanoid pathway synthesizing the building blocks of the lignin polymer. As such, we provide proof of concept of this chemical biology approach to screen for inhibitors of lignification and present a broad array of putative inhibitors of lignin deposition for further characterization.
authors.affiliation: Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87 Umea, Sweden (R.D., E.P.);Compound Screening Facility, VIB, Ghent University, B-9052 Gent, Belgium (D.A., L.N.); andArrhenius Laboratories, Department of Ecology, Environment, and Plant Sciences, Stockholm University, 160 91 Stockholm, Sweden (E.P.).
authors.forename: Dorien
authors.initials: D
authors.lastname: Van de Wouwer
authors.affiliation: Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87 Umea, Sweden (R.D., E.P.);Compound Screening Facility, VIB, Ghent University, B-9052 Gent, Belgium (D.A., L.N.); andArrhenius Laboratories, Department of Ecology, Environment, and Plant Sciences, Stockholm University, 160 91 Stockholm, Sweden (E.P.).
authors.forename: Ruben
authors.identifier: 0000-0001-5848-3138
authors.initials: R
authors.lastname: Vanholme
authors.affiliation: Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87 Umea, Sweden (R.D., E.P.);Compound Screening Facility, VIB, Ghent University, B-9052 Gent, Belgium (D.A., L.N.); andArrhenius Laboratories, Department of Ecology, Environment, and Plant Sciences, Stockholm University, 160 91 Stockholm, Sweden (E.P.).
authors.forename: Raphaël
authors.initials: R
authors.lastname: Decou
authors.affiliation: Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87 Umea, Sweden (R.D., E.P.);Compound Screening Facility, VIB, Ghent University, B-9052 Gent, Belgium (D.A., L.N.); andArrhenius Laboratories, Department of Ecology, Environment, and Plant Sciences, Stockholm University, 160 91 Stockholm, Sweden (E.P.).
authors.forename: Geert
authors.identifier: 0000-0002-0337-2999
authors.initials: G
authors.lastname: Goeminne
authors.affiliation: Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87 Umea, Sweden (R.D., E.P.);Compound Screening Facility, VIB, Ghent University, B-9052 Gent, Belgium (D.A., L.N.); andArrhenius Laboratories, Department of Ecology, Environment, and Plant Sciences, Stockholm University, 160 91 Stockholm, Sweden (E.P.).
authors.forename: Dominique
authors.initials: D
authors.lastname: Audenaert
authors.affiliation: Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87 Umea, Sweden (R.D., E.P.);Compound Screening Facility, VIB, Ghent University, B-9052 Gent, Belgium (D.A., L.N.); andArrhenius Laboratories, Department of Ecology, Environment, and Plant Sciences, Stockholm University, 160 91 Stockholm, Sweden (E.P.).
authors.forename: Long
authors.initials: L
authors.lastname: Nguyen
authors.affiliation: Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87 Umea, Sweden (R.D., E.P.);Compound Screening Facility, VIB, Ghent University, B-9052 Gent, Belgium (D.A., L.N.); andArrhenius Laboratories, Department of Ecology, Environment, and Plant Sciences, Stockholm University, 160 91 Stockholm, Sweden (E.P.).
authors.forename: René
authors.identifier: 0000-0003-0964-1253
authors.initials: R
authors.lastname: Höfer
authors.affiliation: Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87 Umea, Sweden (R.D., E.P.);Compound Screening Facility, VIB, Ghent University, B-9052 Gent, Belgium (D.A., L.N.); andArrhenius Laboratories, Department of Ecology, Environment, and Plant Sciences, Stockholm University, 160 91 Stockholm, Sweden (E.P.).
authors.forename: Edouard
authors.identifier: 0000-0002-6959-3284
authors.initials: E
authors.lastname: Pesquet
authors.affiliation: Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87 Umea, Sweden (R.D., E.P.);Compound Screening Facility, VIB, Ghent University, B-9052 Gent, Belgium (D.A., L.N.); andArrhenius Laboratories, Department of Ecology, Environment, and Plant Sciences, Stockholm University, 160 91 Stockholm, Sweden (E.P.).
authors.forename: Bartel
authors.identifier: 0000-0002-7214-7170
authors.initials: B
authors.lastname: Vanholme
authors.affiliation: Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium (D.V.d.W., R.V., G.G., R.H., B.V., W.B.);Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, 901 87 Umea, Sweden (R.D., E.P.);Compound Screening Facility, VIB, Ghent University, B-9052 Gent, Belgium (D.A., L.N.); andArrhenius Laboratories, Department of Ecology, Environment, and Plant Sciences, Stockholm University, 160 91 Stockholm, Sweden (E.P.) woboe@psb.vib-ugent.be.
authors.forename: Wout
authors.identifier: 0000-0003-1495-510X
authors.initials: W
authors.lastname: Boerjan
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D053898:Biosynthetic Pathways / Q000187:drug effects / Q000235:genetics; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D016000:Cluster Analysis; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000145:classification / Q000494:pharmacology; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D057166:High-Throughput Screening Assays / Q000379:methods; D007463:Iodobenzoates / Q000737:chemistry / Q000494:pharmacology*; D008031:Lignin / Q000378:metabolism*; D013058:Mass Spectrometry; D015394:Molecular Structure; D020005:Propanols / Q000378:metabolism; D036226:Seedlings / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D050564:Trans-Cinnamate 4-Monooxygenase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D007463:Iodobenzoates; D020005:Propanols; C439395:1-phenylpropanol; D008031:Lignin; D050564:Trans-Cinnamate 4-Monooxygenase