Characterization of dehydrodolichyl diphosphate synthase of Arabidopsis thaliana, a key enzyme in dolichol biosynthesis

Abstract

The enzyme dehydrodolichyl diphosphate (dedol-PP) synthase is a cis-prenyltransferase that catalyzes the synthesis of dedol-PP, the long-chain polyprenyl diphosphate used as a precursor for the synthesis of dolichyl phosphate. Here we report the cloning and characterization of a cDNA from Arabidopsis thaliana encoding dedol-PP synthase. The identity of the cloned enzyme was confirmed by functional complementation of a yeast mutant strain defective in dedol-PP synthase activity together with the detection of high levels of dedol-PP synthase activity in the transformed yeast mutant. The A. thaliana dedol-PP synthase mRNA was detected at high levels in roots but was hardly detected in flowers, leaves, stems and in A. thaliana suspension-cultured cells. ß

Comprehensive Tissue-Specific Transcriptome Analysis Reveals Distinct Regulatory Programs during Early Tomato Fruit Development

“…S4). This cluster also contained genes encoding proteins associated with the secretory pathway, including dehydrodolichyl diphosphate synthase, an enzyme that catalyzes the formation of lipids used in the production of N-glycans (Cunillera et al, 2000), and two annexins, potentially involved in exocytosis (Carroll et al, 1998).…”

Monoterpenes in the glandular trichomes of tomato are synthesized from a neryl diphosphate precursor rather than geranyl diphosphate

“…To date, two DDPS genes have been investigated in A. thaliana. At2g23410 was shown to complement a Saccharomyces cereviseae (baker's yeast) rer2 DDPS structural gene mutation, indicating that this A. thaliana protein could catalyze the production of dolichols in yeast (10). At1g11755/lew1 was identified in a mutant screen for plants with a leaf-wilting phenotype under normal growth conditions (25).…”

“…In contrast, the isoprene units of some long-chain plant terpenoids such as rubber and dolichols (the latter group of compounds is present in bacteria, fungi, and animals as well) are linked to each other in the cis (Z) conformation (9,10). They are synthesized from Z-prenyl diphosphates, which are in turn synthesized by cis-prenyltransferases directly from IPP and DMAPP (although in the case of rubber, the starting substrate may be E,E-FPP and IPP) (11).…”

“…Several cis-prenyltransferases have been characterized in bacteria and yeast (13)(14)(15). Bioinformatic analysis of plant sequences identified several proteins with sequence similarity to these cis-prenyltransferases (10,11). The protein encoded by one Arabidopsis thaliana putative cis-prenyltransferase gene catalyzes the formation of C90 to C130 cis-prenyl diphosphates in vitro (10).…”

“…Bioinformatic analysis of plant sequences identified several proteins with sequence similarity to these cis-prenyltransferases (10,11). The protein encoded by one Arabidopsis thaliana putative cis-prenyltransferase gene catalyzes the formation of C90 to C130 cis-prenyl diphosphates in vitro (10). It was hypothesized that this enzyme is involved in the biosynthesis of dolichols in vivo, but no direct in planta evidence was presented.…”