Cloning and characterization of two different types of geranyl diphosphate synthases from Norway spruce (Picea abies).
Geranyl diphosphate (GPP), the universal precursor of monoterpenes, is formed from isopentenyl diphosphate and dimethylallyl diphosphate by the action of geranyl diphosphate synthase, one of the key branchpoint enzymes of terpene biosynthesis. Three types of GPP synthase can be distinguished in plants based on sequence similarity and subunit architecture, but until now individual species have been reported to contain only one of these types. Here we show that the conifer, Norway spruce (Picea abies), contains two different types of GPP synthase belonging to two separate groups of homodimeric proteins. One enzyme, designated PaIDS2 (P. abies isoprenyl diphosphate synthase 2), has high sequence similarity to other gymnosperm GPP synthases. It produces solely GPP in in vitro assays after expression in Escherichia coli and likely participates in monoterpene biosynthesis accompanying induced oleoresin formation, based on dramatic increases in transcript level after methyl jasmonate application. The other enzyme, designated PaIDS3, has highest similarity to the previously reported Arabidopsis thaliana GPP synthase and several other angiosperm sequences, and is not associated with induced oleoresin formation in Norway spruce. In vitro assay of this protein and one encoded by a similar gene sequence from Quercus robur gave substantial amounts of the larger prenyl diphosphates, FPP and GGPP, in addition to GPP. Hence these proteins may not be involved in monoterpene formation and could conceivably form products in addition to GPP in planta.
Max Planck Institute for Chemical Ecology, Department of Biochemistry, Hans-Knöll-Strasse 8, D-07745 Jena, Germany. aschmidt@ice.mpg.de
Axel
A
Schmidt
Jonathan
J
Gershenzon
D000085:Acetates / Q000494:pharmacology; D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003517:Cyclopentanes / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D011756:Diphosphates / Q000378:metabolism*; D004224:Diterpenes / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D008969:Molecular Sequence Data; D054883:Oxylipins / Q000494:pharmacology; D010802:Phylogeny; D028222:Picea / Q000201:enzymology* / Q000235:genetics*; D029963:Quercus / Q000187:drug effects; D013729:Terpenes / Q000378:metabolism
D000085:Acetates; D003517:Cyclopentanes; D018076:DNA, Complementary; D011756:Diphosphates; D004224:Diterpenes; D054883:Oxylipins; D013729:Terpenes; C511282:geranyl diphosphate; C072239:methyl jasmonate; D019883:Alkyl and Aryl Transferases