Department of Genetics, Cell Biology & Development and the Developmental Biology Center, University of Minnesota, Minneapolis, Minnesota, United States of America.
Center for Individualized Medicine, Mayo Clinic College of Medicine, Rochester, Minnesota, United States of America.
PLoS Biol. 2022 May 20;20(5):e3001660. doi: 10.1371/journal.pbio.3001660. eCollection 2022 May.
In polarized epithelial cells, receptor-ligand interactions can be restricted by different spatial distributions of the 2 interacting components, giving rise to an underappreciated layer of regulatory complexity. We explored whether such regulation occurs in the Drosophila wing disc, an epithelial tissue featuring the TGF-β family member Decapentaplegic (Dpp) as a morphogen controlling growth and patterning. Dpp protein has been observed in an extracellular gradient within the columnar cell layer of the disc, but also uniformly in the disc lumen, leading to the question of how graded signaling is achieved in the face of 2 distinctly localized ligand pools. We find the Dpp Type II receptor Punt, but not the Type I receptor Tkv, is enriched at the basolateral membrane and depleted at the junctions and apical surface. Wit, a second Type II receptor, shows a markedly different behavior, with the protein detected on all membrane regions but enriched at the apical side. Mutational studies identified a short juxtamembrane sequence required for basolateral restriction of Punt in both wing discs and mammalian Madin-Darby canine kidney (MDCK) cells. This basolateral targeting (BLT) determinant can dominantly confer basolateral localization on an otherwise apical receptor. Rescue of punt mutants with transgenes altered in the targeting motif showed that flies expressing apicalized Punt due to the lack of a functional BLT displayed developmental defects, female sterility, and significant lethality. We also show that apicalized Punt does not produce an ectopic signal, indicating that the apical pool of Dpp is not a significant signaling source even when presented with Punt. Instead, we find that basolateral presentation of Punt is required for optimal signaling. Finally, we present evidence that the BLT acts through polarized sorting machinery that differs between types of epithelia. This suggests a code whereby each epithelial cell type may differentially traffic common receptors to enable distinctive responses to spatially localized pools of extracellular ligands.
在极化的上皮细胞中,受体-配体相互作用可以受到 2 种相互作用成分的不同空间分布限制,从而产生一种被低估的调控复杂性层次。我们探讨了这种调控是否发生在果蝇翅盘中,这是一种上皮组织,其特征是 TGF-β 家族成员 Decapentaplegic(Dpp)作为一种形态发生素,控制生长和模式形成。Dpp 蛋白已在盘状细胞层的柱状细胞层内观察到作为细胞外梯度,但也均匀地存在于盘状腔中,这就提出了一个问题,即在面对 2 种明显定位于配体池的情况下,如何实现分级信号传递。我们发现 Dpp 型 II 受体 Punt,但不是 I 型受体 Tkv,在基底外侧膜上富集,在连接和顶端表面上耗尽。Wit 是第二种 II 型受体,其行为明显不同,在所有膜区域都检测到蛋白,但在顶端侧富集。突变研究确定了一个短的跨膜序列,该序列对于 Punt 在翅膀和哺乳动物 Madin-Darby 犬肾(MDCK)细胞中的基底外侧限制是必需的。这种基底外侧靶向(BLT)决定因素可以在其他情况下将顶端受体主要定位于基底外侧。用在靶向基序中改变的转基因拯救 punt 突变体表明,由于缺乏功能性 BLT 而表达顶端化 Punt 的苍蝇表现出发育缺陷、雌性不育和显着的致死性。我们还表明,即使存在 Punt,顶端化 Punt 也不会产生异位信号,表明即使存在 Punt,Dpp 的顶端池也不是重要的信号源。相反,我们发现 Punt 的基底外侧呈递对于最佳信号传递是必需的。最后,我们提供了证据表明,BLT 通过上皮细胞类型之间不同的极化分拣机制起作用。这表明了一种编码,即每种上皮细胞类型都可以不同地运输常见受体,从而对空间定位的细胞外配体池产生独特的反应。