Ferulate-5-hydroxylase from Arabidopsis thaliana defines a new family of cytochrome P450-dependent monooxygenases.

The fah1 mutant of Arabidopsis is defective in the accumulation of sinapic acid-derived metabolites, including the guaiacyl-syringyl lignin typical of angiosperms. Earlier results indicated that the FAH1 locus encodes ferulate-5-hydroxylase (F5H), a cytochrome P450-dependent monooxygenase (P450) of the general phenylpropanoid pathway. We have cloned the gene encoding this P450 by T-DNA tagging and have confirmed the identity of the cloned gene by complementation of the mutant phenotype. F5H shows 34% amino acid sequence identity with the avocado ripening-induced P450 CYP71A1 and 32% identity with the flavonoid-3',5'-hydroxylases of Petunia hybrida. In contrast, it shares much less homology with cinnamate-4-hydroxylase, a P450 that catalyzes the hydroxylation of cinnamic acid three steps earlier in the general phenylpropanoid pathway. Since the highest degree of identity between F5H and previously sequenced P450s is only 34%, F5H identifies a new P450 subfamily that has been designated CYP84.

[1]  E. Grill,et al.  A protein phosphatase 2C involved in ABA signal transduction in Arabidopsis thaliana. , 1994, Science.

[2]  S. Yamazaki,et al.  Importance of the proline-rich region following signal-anchor sequence in the formation of correct conformation of microsomal cytochrome P-450s. , 1993, Journal of biochemistry.

[3]  A. Brash,et al.  Molecular cloning of an allene oxide synthase: a cytochrome P450 specialized for the metabolism of fatty acid hydroperoxides. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[4]  The maize Dwarf3 gene encodes a cytochrome P450-mediated early step in Gibberellin biosynthesis. , 1995, The Plant cell.

[5]  R. Last,et al.  Arabidopsis Mutants Lacking Phenolic Sunscreens Exhibit Enhanced Ultraviolet-B Injury and Oxidative Damage , 1995, Plant physiology.

[6]  T. Kutchan,et al.  Molecular cloning and heterologous expression of a cDNA encoding berbamunine synthase, a C--O phenol-coupling cytochrome P450 from the higher plant Berberis stolonifera. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[7]  F. Nagy,et al.  Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter , 1985, Nature.

[8]  R. Sederoff,et al.  Genetic engineering of wood , 1991 .

[9]  M. Van Montagu,et al.  Ti plasmid vector for the introduction of DNA into plant cells without alteration of their normal regeneration capacity , 1983, The EMBO journal.

[10]  Yoshikazu Tanaka,et al.  Cloning and expression of cytochrome P450 genes controlling flower colour , 1993, Nature.

[11]  P. Goldsbrough,et al.  Characterisation of the genes for ribosomal RNA in flax. , 1981, Nucleic acids research.

[12]  H F Kern,et al.  Selection of AUG initiation codons differs in plants and animals. , 1987, The EMBO journal.

[13]  F. Durst,et al.  Plant Cytochromes P450: An Overview , 1995, Drug metabolism and drug interactions.

[14]  D. Ohta,et al.  Molecular cloning and sequencing of a cDNA encoding mung bean cytochrome P450 (P450C4H) possessing cinnamate 4-hydroxylase activity. , 1993, Biochemical and biophysical research communications.

[15]  B. Halkier,et al.  The primary sequence of cytochrome P450tyr, the multifunctional N-hydroxylase catalyzing the conversion of L-tyrosine to p-hydroxyphenylacetaldehyde oxime in the biosynthesis of the cyanogenic glucoside dhurrin in Sorghum bicolor (L.) Moench. , 1995, Archives of biochemistry and biophysics.

[16]  Frans,et al.  Genes Galore: A Summary of Methods for Accessing Results from Large-Scale Partial Sequencing of Anonymous Arabidopsis cDNA Clones , 1994, Plant physiology.

[17]  F. Nagy,et al.  Brassinosteroids Rescue the Deficiency of CYP90, a Cytochrome P450, Controlling Cell Elongation and De-etiolation in Arabidopsis , 1996, Cell.

[18]  N. Lewis,et al.  Lignin: occurrence, biogenesis and biodegradation. , 1990, Annual review of plant physiology and plant molecular biology.

[19]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[20]  H. Yu,et al.  Sequence analysis of ripening-related cytochrome P-450 cDNAs from avocado fruit. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[21]  A. Paterson,et al.  Genome mapping in plants , 1993 .

[22]  Z. Ye,et al.  An alternative methylation pathway in lignin biosynthesis in Zinnia. , 1994, The Plant cell.

[23]  K. Cornish,et al.  The Major Protein of Guayule Rubber Particles Is a Cytochrome P450 , 1995, The Journal of Biological Chemistry.

[24]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[25]  J. Garnier,et al.  Isolation and sequence of a cDNA encoding the Jerusalem artichoke cinnamate 4-hydroxylase, a major plant cytochrome P450 involved in the general phenylpropanoid pathway. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[26]  C. Chapple,et al.  An Arabidopsis mutant defective in the general phenylpropanoid pathway. , 1992, The Plant cell.

[27]  M. J. Coon,et al.  The P450 superfamily: update on new sequences, gene mapping, accession numbers, early trivial names of enzymes, and nomenclature. , 1993, DNA and cell biology.

[28]  J. Giraudat,et al.  RPS2 of Arabidopsis thaliana: a leucine-rich repeat class of plant disease resistance genes. , 1994, Science.