Stewart B A, Mohtashami M, Zhou L, Trimble W S, Boulianne G L
Program in Developmental Biology, The Hospital for Sick Children Research Institute, Toronto, Ontario, M5G 1X8, Canada.
Dev Biol. 2001 Jun 1;234(1):13-23. doi: 10.1006/dbio.2001.0228.
The wing of Drosophila melanogaster has long been used as a model system to characterize intermolecular interactions important in development. Implicit in our understanding of developmental processes is the proper trafficking and sorting of signaling molecules, although the precise mechanisms that regulate membrane trafficking in a developmental context are not well studied. We have therefore chosen the Drosophila wing to assess the importance of SNARE-dependent membrane trafficking during development. N-Ethylmaleimide-sensitive fusion protein (NSF) is a key component of the membrane-trafficking machinery and we constructed a mutant form of NSF whose expression we directed to the developing wing margin. This resulted in a notched-wing phenotype, the severity of which was enhanced when combined with mutants of VAMP/Synaptobrevin or Syntaxin, indicating that it results from impaired membrane trafficking. Importantly, we find that the phenotype is also enhanced by mutations in genes for wingless and components of the Notch signaling pathway, suggesting that these signaling pathways were disrupted. Finally, we used this phenotype to conduct a screen for interacting genes, uncovering two Notch pathway components that had not previously been linked to wing development. We conclude that SNARE-mediated membrane trafficking is an important component of wing margin development and that dosage-sensitive developmental pathways will act as a sensitive reporter of partial membrane-trafficking disruption.
黑腹果蝇的翅膀长期以来一直被用作一个模型系统，以表征在发育过程中重要的分子间相互作用。在我们对发育过程的理解中，隐含着信号分子的正确运输和分选，尽管在发育背景下调节膜运输的精确机制尚未得到充分研究。因此，我们选择了果蝇翅膀来评估SNARE依赖性膜运输在发育过程中的重要性。N - 乙基马来酰亚胺敏感融合蛋白（NSF）是膜运输机制的关键组成部分，我们构建了一种NSF的突变形式，并将其表达定向到发育中的翅膀边缘。这导致了缺刻翅表型，当与VAMP/突触小泡蛋白或Syntaxin的突变体结合时，其严重程度会增强，表明这是由膜运输受损导致的。重要的是，我们发现无翅基因和Notch信号通路成分的突变也会增强这种表型，这表明这些信号通路被破坏了。最后，我们利用这种表型进行了相互作用基因的筛选，发现了两个以前未与翅膀发育相关联的Notch通路成分。我们得出结论，SNARE介导的膜运输是翅膀边缘发育的重要组成部分，并且剂量敏感型发育通路将作为部分膜运输破坏的敏感报告器。
