Small molecules intercept Notch signaling and the early secretory pathway
Small molecules intercept Notch signaling and the early secretory pathway
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DOI：
10.1038/nchembio.1356
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发表时间：
2013-11-01
影响因子：
14.8
通讯作者：
Kaether, Christoph
中科院分区：
文献类型：
--
作者：
Kraemer, Andreas;Mentrup, Torben;Kaether, Christoph
Notch信号传导在许多细胞命运决定中具有关键作用，其异常活性导致发育障碍和癌症。为了鉴定影响Notch信号传导的分子，我们使用自动显微镜筛选了近17，000种化合物，以监测配体独立的Notch增强的GFP（eGFP）报告基因的运输和加工。通过生物化学和细胞测定以及使用斑马鱼在体内进行的体外命中表征导致了五种经验证的化合物，其中四种通过抑制γ-分泌酶诱导报告基因在质膜上的积累。一种化合物，二氢吡啶FLI-06，以不同于布雷菲德菌素A和杀高尔基体剂A的方式破坏高尔基体。FLI-06在离开内质网（ER）之前的一个步骤抑制一般分泌，这伴随着ER的小管到片层形态学转变，使FLI-06成为在分泌运输的早期阶段起作用的第一个小分子。这些数据突出了表型筛选的力量，使中央细胞信号通路的调查。
Notch signaling has a pivotal role in numerous cell-fate decisions, and its aberrant activity leads to developmental disorders and cancer. To identify molecules that influence Notch signaling, we screened nearly 17,000 compounds using automated microscopy to monitor the trafficking and processing of a ligand-independent Notch-enhanced GFP (eGFP) reporter. Characterization of hits in vitro by biochemical and cellular assays and in vivo using zebrafish led to five validated compounds, four of which induced accumulation of the reporter at the plasma membrane by inhibiting gamma-secretase. One compound, the dihydropyridine FLI-06, disrupted the Golgi apparatus in a manner distinct from that of brefeldin A and golgicide A. FLI-06 inhibited general secretion at a step before exit from the endoplasmic reticulum (ER), which was accompanied by a tubule-to-sheet morphological transition of the ER, rendering FLI-06 the first small molecule acting at such an early stage in secretory traffic. These data highlight the power of phenotypic screening to enable investigations of central cellular signaling pathways.