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Research Papers

An eGFP-Based Genetic Screen for Defects in Light-Triggered Subcelluar Translocation of the Drosophila Photoreceptor Channel TRPL

Nina Meyer, Claudia Oberegelsbacher, Thomas D. Dürr, Andrea Schäfer and Armin Huber

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Signaling at the plasma membrane is modulated by up- and down-regulation of signaling proteins. A prominent example for this type of regulation is the Drosophila TRPL ion channel that changes its spatial distribution within the photoreceptor cell. In dark-raised flies TRPL is localized in the rhabdomeral photoreceptor membrane and it translocates to the cell body upon illumination. It has been shown that TRPL translocation depends on the activation of the phototransduction cascade and requires the presence of functional rhodopsin as well as Ca2+-influx through a second light-activated ion channel, TRP. However, little is known about the cell biological mechanism underlying TRPL translocation. Here we describe a FRT/FLP screen designed to isolate mutants defective in TRPL internalization based on the localization of eGFP-tagged TRPL in the eyes of living flies. We mutated chromosome arms 2L, 2R, and 3R and isolated 12 mutants that failed to internalize TRPL. We found that four mutants did not complement genes known to affect TRPL translocation, which are trp, ninaE, and inaD. Two of the isolated mutants represent new alleles of trp and ninaE. The trp allele contains a premature stop codon after amino acid 884, whereas the ninaE allele has a mutation resulting in the substitution P193S. As determined biochemically no TRP or rhodopsin protein, respectively, was expressed in the eyes of these mutants. The absence of TRP or rhodopsin in the isolated mutants readily explains the defect in TRPL internalization and proves the feasibility of our genetic screen.

Authors

Nina Meyer

University of Hohenheim

Claudia Oberegelsbacher

University of Hohenheim

Thomas D. Dürr

University of Hohenheim

Andrea Schäfer

University of Hohenheim

Armin Huber

University of Hohenheim

Purchase article for $19

Subscribe to this journal for $79/year