CYP86A1 fromArabidopsis thalianaEncodes a Cytochrome P450-Dependent Fatty Acid Omega-Hydroxylase

Benveniste, Irène; Tijet, Nathalie; Adas, Fadi; Philipps, Gabriel; Salaün, Jean‐Pierre; Durst, Francis

doi:10.1006/bbrc.1998.8156

Cited by 127 publications

(86 citation statements)

References 30 publications

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“…Also, the recent transcriptome analysis of cork phelloderm (16) identified homologs with close similarity to CYP86A1, rather than other members of this P450 subfamily, supporting the association of CYP86A1 with suberin biosynthesis. In vitro fatty acid -oxidase activity has also been detected for recombinant CYP86A1 expressed in yeast (31). The importance of CYP86A1 for accumulation of oxidized fatty acids in suberin was confirmed by the chemical analysis of suberin in roots that showed a 50-75% reduction in several -oxidized monomers including 16-and 18-carbon dicarboxylic acids (SI Fig.…”

Section: Cooverexpression Of Gpat5 and Cyp86a1 Produces Typical Suberinmentioning

confidence: 81%

Identification of acyltransferases required for cutin biosynthesis and production of cutin with suberin-like monomers

Li¹,

Beisson²,

Koo

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

351

342

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Cutin and suberin are the two major lipid-based polymers of plants. Cutin is the structural polymer of the epidermal cuticle, the waterproof layer covering primary aerial organs and which is often the structure first encountered by phytopathogens. Suberin contributes to the control of diffusion of water and solutes across internal root tissues and in periderms. The enzymes responsible for assembly of the cutin polymer are largely unknown. We have identified two Arabidopsis acyltransferases essential for cutin biosynthesis, glycerol-3-phosphate acyltransferase (GPAT) 4 and GPAT8. Double knockouts gpat4/gpat8 were strongly reduced in cutin and were less resistant to desiccation and to infection by the fungus Alternaria brassicicola. They also showed striking defects in stomata structure including a lack of cuticular ledges between guard cells, highlighting the importance of cutin in stomatal biology. Overexpression of GPAT4 or GPAT8 in Arabidopsis increased the content of C16 and C18 cutin monomers in leaves and stems by 80%. In order to modify cutin composition, the acyltransferase GPAT5 and the cytochrome P450-dependent fatty acyl oxidase CYP86A1, two enzymes associated with suberin biosynthesis, were overexpressed. When both enzymes were overexpressed together the epidermal polyesters accumulated new C20 and C22 -hydroxyacids and ␣, -diacids typical of suberin, and the fine structure and water-barrier function of the cuticle were altered. These results identify GPATs as partners of fatty acyl oxidases in lipid polyester synthesis and indicate that their cooverexpression provides a strategy to probe the role of cutin composition and quantity in the function of plant cuticles. P lant surface lipids fulfill critical roles in the control of water and gas exchange, as protection from pathogens and UV radiation, as structural components, and to prevent cell fusions during organogenesis (1-3). These lipids are organized into the cuticle, a complex hydrophobic layer that covers the epidermis of plant leaves and other aerial organs and therefore is one of the largest biological interfaces in nature. The framework of the cuticle layer is provided by cutin, a plant-specific polyester composed of omega-substituted fatty acids and glycerol monomers (1, 4, 5). This insoluble polyester matrix is embedded and covered with waxes, a mixture of fatty acid derivatives that is easily extractable in organic solvents and has thus been more amenable to study than cutin (2, 6). Suberin is another type of cell-wall-associated lipid polymer, the most well known form being cork. It is composed of aliphatic and aromatic domains and is found in roots, the periderm of stems, and other tissues where it functions to restrict movement of water and ions across cell walls (1,7,8). Suberin also differs from cutin in that it is usually deposited abutting the inner face of the primary cell wall, whereas cutin is deposited at the outer face.Although cutin, one of the most abundant lipid polymers of nature, is the structural polymer of the plant cutic...

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Section: Cooverexpression Of Gpat5 and Cyp86a1 Produces Typical Suberinmentioning

confidence: 81%

Identification of acyltransferases required for cutin biosynthesis and production of cutin with suberin-like monomers

Li¹,

Beisson²,

Koo

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

351

342

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“…With linoleic acid as a substrate, there is no change in the absorbance between 210 and 350 nm including absorption at 234 nm corresponding to the formation of a conjugated diene hydroperoxy moiety during the reaction. 2 This finding indicated that rice ␣-oxygenase could not abstract the bisallylic proton to form conjugated diene hydroperoxide but exclusively abstract ␣-proton.…”

Section: Resultsmentioning

confidence: 98%

“…It was confirmed that the enzymes localize on the mitochondria and peroxisomes. Cytochrome P450s in the endoplasmic reticulum catalyze the fatty acid -hydroxylation (2). There are at least three types of ␣-oxidation systems.…”

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confidence: 99%

Catalytic Properties of Rice α-Oxygenase

Koeduka

Matsui

Akakabe

et al. 2002

Journal of Biological Chemistry

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Long-chain fatty acids can be metabolized to C n؊1 aldehydes by ␣-oxidation in plants. The reaction mechanism of the enzyme has not been elucidated. In this study, a complete nucleotide sequence of fatty acid ␣-oxygenase gene in rice plants (Oryza sativa) was isolated. The deduced amino acid sequence showed some similarity with those of mammalian prostaglandin H synthases (PGHSs). The gene was expressed in Escherichia coli and purified to apparently homogenous state. It showed the highest activity with linoleic acid and predominantly formed 2-hydroperoxide of the fatty acid (C n ), which is then spontaneously decarboxylated to form corresponding C n؊1 aldehyde. With linoleic or linoleic acids as a substrate, rice ␣-oxygenase formed no product having a max at approximately 234 nm, which indicated that the enzyme could not oxygenize the pentadiene system in the substrate. The spectroscopic feature of the purified enzyme in its ferrous state is similar to that of mammalian PGHS, whereas that of dithionite-reduced state showed significant difference. Site-directed mutagenesis revealed that His-158, Tyr-380, and Ser-558 were essential for the ␣-oxygenase activity. These residues are conserved in PGHS and known as a heme ligand, a source of a radical species to initiate oxygenation reaction and a residue involved in substrate binding, respectively. This finding suggested that the initial step of the oxygenation reaction in ␣-oxygenase has a high similarity with that of PGHS. The rice ␣-oxygenase activity was inhibited by imidazole but hardly inhibited by nonsteroidal anti-inflammatory drugs, such as aspirin, ibuprofen, and flurbiprofen, which are known as typical PGHS inhibitors. In addition, peroxidase activity could not be detected with ␣-oxygenase when palmitic acid 2-hydroperoxide was used as a substrate. From these findings, the catalytic resemblance between ␣-oxygenase and PGHS seems to be evident, although there still are differences in their substrate recognitions and peroxidation activities.

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“…All members of the CYP86A subfamily in Arabidopsis analyzed so far have a demonstrated fatty acid v-hydroxylase activity in vitro (Benveniste et al, 1998;Rupasinghe et al, 2007). A fatty acid v-hydroxylase activity may also be expected for potato CYP86A33 based on the changes that its deficiency produces in the aliphatic suberin composition that mainly concern the v-functionalized monomers.…”

Section: Significance Of V-functional Fatty Acids In Suberin Biosynthmentioning

confidence: 97%

“…All Arabidopsis proteins included in the study demonstrated in vitro v-hydroxylase activity (Benveniste et al, 1998;Kandel et al, 2007;Rupasinghe et al, 2007) and alsofor CYP94C1-a capacity to yield a,v-diacids (Kandel et al, 2007). A multiple amino acid sequence alignment generated by ClustalW was used to generate two unrooted phylogenetic trees using, respectively, the neighbor-joining and the unweighted pair group method with arithmetic mean algorithms.…”

Section: Cloning and Phylogenetic Analysis Of Potato Cyp86a33 And Cormentioning

confidence: 99%

CYP86A33-Targeted Gene Silencing in Potato Tuber Alters Suberin Composition, Distorts Suberin Lamellae, and Impairs the Periderm's Water Barrier Function

et al. 2008

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Suberin is a cell wall lipid polyester found in the cork cells of the periderm offering protection against dehydration and pathogens. Its biosynthesis and assembly, as well as its contribution to the sealing properties of the periderm, are still poorly understood. Here, we report on the isolation of the coding sequence CYP86A33 and the molecular and physiological function of this gene in potato (Solanum tuberosum) tuber periderm. CYP86A33 was down-regulated in potato plants by RNA interference-mediated silencing. Periderm from CYP86A33-silenced plants revealed a 60% decrease in its aliphatic suberin load and greatly reduced levels of C18:1 v-hydroxyacid (approximately 70%) and a,v-diacid (approximately 90%) monomers in comparison with wild type. Moreover, the glycerol esterified to suberin was reduced by 60% in the silenced plants. The typical regular ultrastructure of suberin, consisting of dark and light lamellae, disappeared and the thickness of the suberin layer was clearly reduced. In addition, the water permeability of the periderm isolated from CYP86A33-silenced lines was 3.5 times higher than that of the wild type. Thus, our data provide convincing evidence for the involvement of v-functional fatty acids in establishing suberin structure and function.

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CYP86A1 fromArabidopsis thalianaEncodes a Cytochrome P450-Dependent Fatty Acid Omega-Hydroxylase

Cited by 127 publications

References 30 publications

Identification of acyltransferases required for cutin biosynthesis and production of cutin with suberin-like monomers

Identification of acyltransferases required for cutin biosynthesis and production of cutin with suberin-like monomers

Catalytic Properties of Rice α-Oxygenase

CYP86A33-Targeted Gene Silencing in Potato Tuber Alters Suberin Composition, Distorts Suberin Lamellae, and Impairs the Periderm's Water Barrier Function

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