title: Random sequencing of an induced Taxus cell cDNA library for identification of clones involved in Taxol biosynthesis.
abstract: Biosynthesis of the anticancer drug Taxol involves 19 enzymatic steps from the universal diterpenoid progenitor geranylgeranyl diphosphate derived by the plastidial methylerythritol phosphate pathway for isoprenoid precursor supply. To gain further insight about Taxol biosynthesis relevant to the improved production of this drug and to draw inferences about the organization, regulation, and origins of this complex natural product pathway, random sequencing of a cDNA library derived from Taxus cuspidata cells (induced for taxoid biosynthesis with methyl jasmonate) was undertaken. This effort revealed surprisingly high abundances for transcripts of several of the 12 defined genes of Taxol biosynthesis, yielded cDNAs encoding two previously uncharacterized cytochrome P450 taxoid hydroxylases, and provided candidate genes for all but one of the remaining seven steps of this extended sequence of reactions.
authors.affiliation: Institute of Biological Chemistry, Washington State University, Pullman, WA 99164-6340, USA.
authors.forename: Stefan
authors.initials: S
authors.lastname: Jennewein
authors.forename: Mark R
authors.initials: MR
authors.lastname: Wildung
authors.forename: MyDoanh
authors.initials: M
authors.lastname: Chau
authors.forename: Kevin
authors.initials: K
authors.lastname: Walker
authors.forename: Rodney
authors.initials: R
authors.lastname: Croteau
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000235:genetics; D000595:Amino Acid Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D004896:Erythritol / Q000031:analogs & derivatives* / Q000378:metabolism; D015723:Gene Library; D008969:Molecular Sequence Data; D017239:Paclitaxel / Q000096:biosynthesis*; D010802:Phylogeny; D016415:Sequence Alignment; D013403:Sugar Phosphates / Q000378:metabolism; D043823:Taxoids / Q000378:metabolism; D020946:Taxus / Q000166:cytology / Q000235:genetics*
chemical_list: C114232:2-C-methylerythritol 4-phosphate; D018076:DNA, Complementary; D018744:DNA, Plant; D013403:Sugar Phosphates; D043823:Taxoids; D003577:Cytochrome P-450 Enzyme System; D019883:Alkyl and Aryl Transferases; D017239:Paclitaxel; D004896:Erythritol