Amarnath Smita, Stevens Leslie M, Stein David S
Department of Molecular Biosciences and Institute for Cellular and Molecular Biology, The University of Texas at Austin, Patterson Labs 532, 2401 Speedway, Austin, TX 78712, USA.
Department of Molecular Biosciences and Institute for Cellular and Molecular Biology, The University of Texas at Austin, Patterson Labs 532, 2401 Speedway, Austin, TX 78712, USA
Development. 2017 Feb 15;144(4):677-686. doi: 10.1242/dev.146076. Epub 2017 Jan 13.
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.
胚胎两端的形成受受体酪氨酸激酶Torso的局部激活调控。Torso及其假定的配体Trunk在早期胚胎中均呈均匀表达。Torso的极化激活需要Torso-like,它由发育中卵母细胞两端相邻的卵泡细胞表达。我们发现,在培养细胞中高水平表达的Torso可被单独施加的Trunk、Torso-like或另一种已知的Torso配体促前胸腺激素激活。除了检测下游信号活性外,还使用双分子荧光互补检测Torso二聚化。当与Torso共转染以及在条件培养基中呈现给表达Torso的细胞时,Trunk和Torso-like具有活性。Trunk和Torso-like也可从条件培养基中被表达Torso的细胞特异性摄取。在与胚胎中相似的低水平Torso情况下,单独的Trunk和Torso-like无效,但协同作用以刺激Torso信号传导。我们的结果表明,Torso与Trunk和Torso-like均相互作用,它们共同协作介导胚胎两端Torso的二聚化和激活。