A female-biased expressed elongase involved in long-chain hydrocarbon biosynthesis and courtship behavior in Drosophila melanogaster.

Drosophila melanogaster produces sexually dimorphic cuticular pheromones that are a key component of the courtship behavior leading to copulation. These molecules are hydrocarbons, with lengths of 23 and 25 carbons in males (mainly with one double bond) and 27 and 29 carbons in females (mainly with two double bonds). Here, we describe an elongase gene, eloF, with female-biased expression. The 771-bp ORF encodes a 257-aa protein that shows the highest sequence identity with mouse SSC1 elongase (33%). The activity of the cDNA expressed in yeast was elongation of saturated and unsaturated fatty acids up to C30. RNAi knockdown in Drosophila led to a dramatic modification of female hydrocarbons, with decreased C29 dienes and increased C25 dienes accompanied by a modification of several courtship parameters: an increase in copulation latency and a decrease in both copulation attempts and copulation. Feminization of the hydrocarbon profile in males by using targeted expression of the transformer gene resulted in high expression levels of eloF, suggesting that the gene is under the control of the sex-determination hierarchy. There is no expression of eloF in Drosophila simulans, which synthesize only C23 and C25 hydrocarbons. These results strongly support the hypothesis that eloF is a crucial enzyme for female pheromone biosynthesis and courtship behavior in D. melanogaster.

A new elongase selectively expressed in Drosophila male reproductive system.

We have identified an elongase gene, elo68alpha, which is specifically transcribed in males. We have characterized the elo68alpha open reading frame, expressed it in fasDelta elo1Delta yeast and showed that it could elongate myristoleic and palmitoleic acids, therefore sharing an Elo1 specificity. This elongase was found to be exclusively expressed in male genital system (testis and ejaculatory bulb). Northern blot analysis showed that the elo68alpha gene was inducible at low temperatures. One P-strain mutant for elo68alpha and three excision lines for this P-element were subsequently studied. The excision line with only 1% elo68alpha expression showed decreased levels of vaccenyl acetate, a male pheromone produced in the ejaculatory bulb. The induction of elo68alpha expression at 21 degrees C was also paralleled with higher vaccenyl acetate production. These results strongly suggest that elo68alpha is involved in the elongation of short unsaturated fatty acids in males and might play a role in vaccenyl acetate biosynthesis.

Mutations in the desat1 gene reduces the production of courtship stimulatory pheromones through a marked effect on fatty acids in Drosophila melanogaster.

In Drosophila melanogaster, desat1 is involved in the synthesis of fatty acids (FAs), some of which are precursors in the production of unsaturated hydrocarbons (HCs) in position 7 (7-HC) that play an important role in mating behaviour. Three GS lines with P-element insertion in the desat1 promoter showed more or less decrease in 7-HC, depending on the site of insertion. The forced transcription of genomic 5'P-flanking sequence led to opposite effects upon 7-HC, depending on the orientation of the insertions. Homozygous GS12251 flies showed particularly low 7-HC levels and severely affected courtship parameters (courtship latency doubled, number of copulation attempts decreased by half). After transposon excision, the HC phenotype was reversed in most lines, showing that the location of the transposon was responsible for the mutant phenotype. In homozygous GS12251 flies, the amounts of FAs and desat1 transcripts were reduced by half, compared to the amounts in heterozygous or wild-type flies. Relative proportions among FAs were quite similar to those of wild-type, with the exception of a slight decrease in myristoleic, palmitoleic and vaccenic acid. As the reduction of desat1 activity in the mutant resulted in a large decrease in both unsaturated and saturated FAs, it could impair FA and lipid metabolism, as it is known in vertebrates.

Involvement of desat1 gene in the control of Drosophila melanogaster pheromone biosynthesis.

Cuticular pheromones in Drosophila melanogaster are unsaturated hydrocarbons with at least one double bond in position 7: 7-tricosene and 7-pentacosene in males and 7,11 -heptacosadiene and 7,11 -nonacosadiene in females. We have previously shown that a desaturase gene, desat1, located in chromosome region 87 C could be involved in this process: the Desat1 enzyme preferentially leads to the synthesis of palmitoleic acid, a precursor of omega7 fatty acids and 7-unsaturated hydrocarbons. Therefore, we have searched for P-elements in the 87 region and mapped them. One was found inserted into the first intron of the desat1 gene. Flies heterozygous for this insertion showed a large decrease in the level of 7-unsaturated hydrocarbons, comparable to that observed in flies heterozygous for a deficiency overlapping desat1. Less than 1 % of flies homozygous for this insertion were viable. They were characterized by dramatic pheromone decreases. After excision of the transposon, the pheromone phenotype was reversed in 69% of the lines and the other excision lines had more or less decreased amounts of 7-unsaturated hydrocarbons. All these results implicate desat1 in the synthesis of Drosophila pheromones.