Reconstitution of Torso signaling in cultured cells suggests a role for both Trunk and Torso-like in receptor activation

• Published in: Development (Cambridge) Vol. 144; no. 4; pp. 677 - 686
• Main Authors: Amarnath, Smita, Stevens, Leslie M, Stein, David S
• Format: Journal Article
• Language: English
• Published: England The Company of Biologists Ltd 15.02.2017
• Subjects: Animals; Cells, Cultured; Culture Media; Dimerization; Drosophila; Drosophila melanogaster - embryology; Drosophila Proteins - metabolism; Embryos; Female; Gene Expression Regulation, Developmental; Green Fluorescent Proteins - metabolism; Insect Hormones - metabolism; Insects; Kinases; Ligands; Ovarian Follicle - metabolism; Protein Multimerization; Protein-tyrosine kinase receptors; Receptor mechanisms; Receptor Protein-Tyrosine Kinases - metabolism; RNA Interference; Signal Transduction; RTK; Receptor tyrosine kinase; Membrane attack complex perforin; Terminal; Drosophila; MACPF
• ISSN: 0950-1991; 1477-9129
• DOI: 10.1242/dev.146076

Formation of the embryonic termini is controlled by the localized activation of the receptor tyrosine kinase Torso. Both Torso and Torso's presumed ligand, Trunk, are expressed uniformly in the early embryo. Polar activation of Torso requires Torso-like, which is expressed by follicle cells adjacent to the ends of the developing oocyte. We find that Torso expressed at high levels in cultured cells is activated by individual application of Trunk, Torso-like or another known Torso ligand, Prothoracicotropic Hormone. In addition to assays of downstream signaling activity, Torso dimerization was detected using bimolecular fluorescence complementation. Trunk and Torso-like were active when co-transfected with Torso and when presented to Torso-expressing cells in conditioned medium. Trunk and Torso-like were also taken up from conditioned medium specifically by cells expressing Torso. At low levels of Torso, similar to those present in the embryo, Trunk and Torso-like alone were ineffective but acted synergistically to stimulate Torso signaling. Our results suggest that Torso interacts with both Trunk and Torso-like, which cooperate to mediate dimerization and activation of Torso at the ends of the embryo.