High-content screening identifies inhibitors of the nuclear translocation of ATF6

作者：Liu, Chun-Lei; Li, Xin; Gan, Lu; He, Yun-Yun; Wang, Li-Li^*; He, Kun-Lun^*

来源：International Journal of Molecular Medicine, 2016, 37(2): 407-414.

DOI：10.3892/ijmm.2015.2442

• high-content screening
• nuclear translocation assay
• activating transcription factor 6
• endoplasmic reticulum stress

Activating transcription factor 6 (ATF6) is a transmembrane protein that consists of a cytoplasmic domain and an endoplasmic reticulum (ER) luminal domain. As unfolded protein levels arise in the ER, the ER cytoplasmic domain of ATF6 moves to the nucleus, where it activates the transcription of a range of genes, including those involved in apoptosis. As ATF6 only becomes functional once it has moved to the nucleus, compounds that inhibit its re-localization are of therapeutic interest. The aim of the present study was to rapidly and accurately identify such compounds using a novel image-based, high-content screening (HCS) technique. The results from the HCS analysis were then confirmed by luciferase reporter assays, western blot analysis and the measurement of cell viability. We found that HCS identified compounds which inhibited ATF6 nuclear translocation with high specificity, as confirmed by the luciferase reporter assay and western blot analysis. Moreover, we demonstrated that 3 of the 80 identified compounds impaired ATF6-mediated induced cell death. The data from this study support the theory that HCS is a novel, high throughput method which can be used for accurate and rapid compound screening.

• 出版日期2016-2
• 单位中国人民解放军总医院