Cloning and functional expression of a cDNA encoding a pheromone gland-specific acyl-CoA Δ 11 -desaturase of the cabbage looper moth, Trichoplusia ni
| Autor: | Weitian Liu, Juliet D. Tang, Stuart J. Miller, Peter W.K. Ma, Douglas C. Knipple, Wendell L. Roelofs, Claire-Lise Rosenfield |
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| Rok vydání: | 1998 |
| Předmět: |
Fatty Acid Desaturases
DNA Complementary Auxotrophy Molecular Sequence Data Saccharomyces cerevisiae Genes Insect Moths Pheromones Open Reading Frames Acyl-CoA chemistry.chemical_compound Cabbage looper Endocrine Glands Complementary DNA Trichoplusia Animals Amino Acid Sequence Cloning Molecular Conserved Sequence Genetics chemistry.chemical_classification Multidisciplinary Sequence Homology Amino Acid biology Fatty acid Biological Sciences biology.organism_classification Yeast Rats Molecular Weight Biochemistry chemistry Sequence Alignment |
| Zdroj: | Proceedings of the National Academy of Sciences. 95:15287-15292 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.95.26.15287 |
| Popis: | 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 Δ 9 -desaturases of animals and fungi, suggesting a common ancestral origin. Unlike metabolic acyl-CoA Δ 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 Δ 11 Z-desaturation mechanism. The largest ORF of the ≈1,250-bp cDNA encodes a 349-aa apoprotein (PDesat-Tn Δ 11 Z) with a predicted molecular mass of 40,240 Da. Its hydrophobicity profile is similar overall to those of rat and yeast Δ 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 Δ 9 Z-desaturases of rat and yeast, respectively. Northern blot analysis revealed an ≈1,250-nt PDesat-Tn Δ 11 Z mRNA that is consistent with the spatial and temporal distribution of Δ 11 -desaturase enzyme activity. Genetic transformation of a desaturase-deficient strain of the yeast Saccharomyces cerevisiae with an expression plasmid encoding PDesat-Tn Δ 11 Z resulted in complementation of the strain’s fatty acid auxotrophy and the production of Δ 11 Z-unsaturated fatty acids. |
| Databáze: | OpenAIRE |
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