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45. Cloning and functional expression of a cDNA encoding a pheromone gland-specific acyl-CoA delta-(super 11)-desaturase of the cabbage looper moth Trichoplusia ni

Author

Knipple, Douglas C., Rosenfield, Claire-Lise, Miller, Stuart J., Liu, Weitian, Tang, Juliet, Ma, Peter W.K., and Roelofs, Wendell L.

Subjects
Cabbage looper -- Genetic aspects, DNA -- Research, Fatty acid desaturases -- Research, Cloning -- Research, Pheromones -- Research, Science and technology
Abstract

Desaturation of coenzyme-A esters of saturated fatty acids is a common feature of sex pheromone biosynthetic pathways in the Lepidoptera. The enzymes that catalyze this step share several biochemical properties with the ubiquitous acyl-CoA [[Delta].sup.9]-desaturases of animals and fungi, suggesting a common ancestral origin. Unlike metabolic acylCoA [[Delta].sup.9]-desaturases, pheromone desaturases have evolved unusual regio- and stereoselective activities that contribute to the remarkable diversity of chemical structures used as pheromones in this large taxonomic group. In this report, we describe the isolation of a cDNA encoding a pheromone gland desaturase from the cabbage looper moth, Trichoplusia ni, a species in which all unsaturated pheromone products are produced via a [[Delta].sup.11]Z-desaturation mechanism. The largest ORF of the [approximately equal to]1,250-bp cDNA encodes a 349-aa apoprotein (PDesat-Tn [[Delta].sup.11]Z) with a predicted molecular mass of 40,240 Da. Its hydrophobicity profile is similar overall to those of rat and yeast [[Delta].sup.9]-desaturases, suggesting conserved transmembrane topology. A 182-aa core domain delimited by conserved histidine-rich motifs implicated in iron-binding and catalysis has 72 and 58% similarity (including conservative substitutions) to acyl-CoA [[Delta].sup.9]Z-desaturases of rat and yeast, respectively. Northern blot analysis revealed an [approximately equal to]1,250-nt PDesat-Tn [[Delta].sup.11]Z mRNA that is consistent with the spatial and temporal distribution of [[Delta]11]-desaturase enzyme activity. Genetic transformation of a desaturase-deficient strain of the yeast Saccharomyces cerevisiae with an expression plasmid encoding PDesat-Tn [[Delta].sup.11]Z resulted in complementation of the strain's fatty acid auxotrophy and the production of [[Delta].sup.11]Z-unsaturated fatty acids.

Published
1998

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