title: High-content screening identifies inhibitors of the nuclear translocation of ATF6.
abstract: 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.
authors.affiliation: Department of Geriatric Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Chun-Lei
authors.initials: CL
authors.lastname: Liu
authors.affiliation: Department of Geriatric Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Geriatric Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Lu
authors.initials: L
authors.lastname: Gan
authors.affiliation: Department of Geriatric Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Yun-Yun
authors.initials: YY
authors.lastname: He
authors.affiliation: Pharmacy Institute of Military Medical Sciences, Beijing 100850, P.R. China.
authors.forename: Li-Li
authors.initials: LL
authors.lastname: Wang
authors.affiliation: Department of Geriatric Cardiology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Kun-Lun
authors.initials: KL
authors.lastname: He
mesh_terms: D051702:Activating Transcription Factor 6 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects* / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D017510:Protein Folding / Q000187:drug effects; D021381:Protein Transport / Q000235:genetics
chemical_list: C497111:ATF6 protein, human; D051702:Activating Transcription Factor 6; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins