Disrupting Hedgehog Cardin–Weintraub sequence and positioning changes cellular differentiation and compartmentalization in vivo
Disrupting Hedgehog Cardin–Weintraub sequence and positioning changes cellular differentiation and compartmentalization in vivo
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DOI：
10.1242/dev.167221
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发表时间：
2018-09
期刊：
影响因子：
4.6
通讯作者：
Philipp Kastl;D. Manikowski;Georg Steffes;S. Schürmann;Shyam Bandari;C. Klämbt;K. Grobe
中科院分区：
文献类型：
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作者：
Philipp Kastl;D. Manikowski;Georg Steffes;S. Schürmann;Shyam Bandari;C. Klämbt;K. Grobe
后生动物刺猬（Hh）形态因子是远离其来源的生长和模式的重要调节因子，尽管它们是作为n端棕榈酰化和c端胆固醇化蛋白产生的，这将它们牢牢地束缚在产生细胞的外质膜小叶上，并限制了它们的传播。克服这一限制的一种机制是两种脂化末端肽的蛋白水解处理，称为脱落，但有效Hh脱落的分子靶位要求仍不明确。在这项工作中，我们以黑腹果蝇为模型，发现n端cardina - weintraub （CW）基序的突变在不同程度上使重组Hh蛋白失活，如果在同一隔室中过表达，则将其转化为内源性Hh功能的抑制因子。在体内，n -棕榈酸盐膜锚的额外去除在很大程度上恢复了内源性Hh功能，支持了蛋白水解CW加工控制Hh增溶的假设。重要的是，我们还观察到连续波重定位损害了三龄翼盘前/后隔室边界的维持。这表明Hh脱落不仅控制了前部细胞的分化，而且还维持了这些接收细胞和后部Hh产生细胞之间的明显物理隔离。摘要：在黑腹果蝇体内，刺猬蛋白脱落（Hedgehog morphogens的蛋白水解裂解）是组织模式和前后室边界维持所必需的。
ABSTRACT Metazoan Hedgehog (Hh) morphogens are essential regulators of growth and patterning at significant distances from their source, despite being produced as N-terminally palmitoylated and C-terminally cholesteroylated proteins, which firmly tethers them to the outer plasma membrane leaflet of producing cells and limits their spread. One mechanism to overcome this limitation is proteolytic processing of both lipidated terminal peptides, called shedding, but molecular target site requirements for effective Hh shedding remained undefined. In this work, by using Drosophila melanogaster as a model, we show that mutagenesis of the N-terminal Cardin–Weintraub (CW) motif inactivates recombinant Hh proteins to variable degrees and, if overexpressed in the same compartment, converts them into suppressors of endogenous Hh function. In vivo, additional removal of N-palmitate membrane anchors largely restored endogenous Hh function, supporting the hypothesis that proteolytic CW processing controls Hh solubilization. Importantly, we also observed that CW repositioning impairs anterior/posterior compartmental boundary maintenance in the third instar wing disc. This demonstrates that Hh shedding not only controls the differentiation of anterior cells, but also maintains the sharp physical segregation between these receiving cells and posterior Hh-producing cells. Summary: Hedgehog shedding, the proteolytic cleavage of Hedgehog morphogens, is required for tissue patterning and anterior/posterior compartment boundary maintenance in vivo in Drosophila melanogaster.