title: A versatile ΦC31 based reporter system for measuring AP-1 and Nrf2 signaling in Drosophila and in tissue culture.
abstract: This paper describes the construction and characterization of a system of transcriptional reporter genes for monitoring the activity of signaling pathways and gene regulation mechanisms in intact Drosophila, dissected tissues or cultured cells. Transgenic integration of the reporters into the Drosophila germline was performed in a site-directed manner, using ΦC31 integrase. This strategy avoids variable position effects and assures low base level activity and high signal responsiveness. Defined integration sites furthermore enable the experimenter to compare the activity of different reporters in one organism. The reporter constructs have a modular design to facilitate the combination of promoter elements (synthetic transcription factor binding sites or natural regulatory sequences), reporter genes (eGFP, or DsRed.T4), and genomic integration sites. The system was used to analyze and compare the activity and signal response profiles of two stress inducible transcription factors, AP-1 and Nrf2. To complement the transgenic reporter fly lines, tissue culture assays were developed in which the same synthetic ARE and TRE elements control the expression of firefly luciferase.
authors.affiliation: Department of Biomedical Genetics, University of Rochester Medical Center, Rochester, New York, United States of America.
authors.forename: Nirmalya
authors.initials: N
authors.lastname: Chatterjee
authors.forename: Dirk
authors.initials: D
authors.lastname: Bohmann
mesh_terms: D000818:Animals; D001435:Bacteriophages / Q000235:genetics*; D001665:Binding Sites; D018929:Cell Culture Techniques; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000166:cytology* / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D015398:Signal Transduction; D013312:Stress, Physiological; D046509:Tissue Culture Techniques; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D029721:Drosophila Proteins; D051267:NF-E2-Related Factor 2; D018808:Transcription Factor AP-1; D048031:JNK Mitogen-Activated Protein Kinases