Functional expression of a locust visual pigment in transgenic Drosophila melanogaster.
The cDNA encoding a visual pigment of the locust Schistocerca gregaria has been inserted into the germline of the ninaE mutant of Drosophila melanogaster by P-element-mediated transformation. Functional expression has been documented by recording light-regulated electroretinograms in transgenic flies. The spectral properties of the expressed visual pigment were determined with detergent-solubilized material, prepared from the eyecups of the transgenic D. melanogaster. The recombinant locust pigment, as well as the genuine pigment of the fruitfly (Rh1) that served as a control for transformation/expression, showed photoreversibility between the pigment and metapigment forms. The absorptions of the difference spectra identify the locust visual pigment as a short wavelength-absorbing, blue-light-sensitive photoreceptor. The absorption maxima are similar to those recorded on living locust animals. These results show that, although locust visual pigments contain 11-cis retinal as chromophore, the expressed protein is able to adopt 3-hydroxyretinal that is provided by the transgenic fruitflies. The electrophysiological recordings reveal that the locust visual pigment is able to induce phototransduction in the fruitfly. The reported results have two important consequences: On the one hand, the binding site of the locust opsin is apparently able to interact with the 3-hydroxyretinal from Drosophila in a way that the biological signal generated by the photoisomerization of the chromophore can be used by the protein to adopt a physiologically active conformation. On the other hand, despite the relatively large phylogenetic distance between both insect species, the extent of conservation between the protein domains thought to be involved in G-protein activation is striking.
Engels
A
A
Max-Planck-Institut für Strahlenchemie, Mülheim an der Ruhr, Germany.
Reichert
H
H
Gehring
W J
WJ
Gärtner
W
W
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary; D004072:Digitonin / Q000737:chemistry; D004331:Drosophila melanogaster / Q000235:genetics*; D005822:Genetic Vectors; D011994:Recombinant Proteins / Q000235:genetics; D012168:Retinal Pigments / Q000235:genetics*; D014170:Transformation, Genetic
D017931:DNA Primers; D018076:DNA, Complementary; D011994:Recombinant Proteins; D012168:Retinal Pigments; D004072:Digitonin