期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 154, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/60652
关键词
Medicine; Issue 154; hepatitis B virus; drug screening; high-throughput; virus-host interaction; Smc5/6; protein-protein interaction; nitazoxanide
资金
- Ministry of Education, Culture, Sports, Science, and Technology, Japan [19H03430, 17K09405, 19J11829]
- Japan Agency for Medical Research and Development, AMED [JP19fk021005]
- AMED [JP19fk0310102]
- Japan Foundation for Applied Enzymology
- Kobayashi Foundation for Cancer Research
- GSK Japan Research Grant 2018
- Miyakawa Memorial Research Foundation
- [18H05024]
- Grants-in-Aid for Scientific Research [19H03430, 19J11829, 17K09405] Funding Source: KAKEN
There is an urgent need for novel therapeutic agents for hepatitis B virus (HBV) infection. Although currently available nucleos(t)ide analogs potently inhibit viral replication, they have no direct effect on the expression of viral proteins transcribed from a viral covalently closed circular DNA (cccDNA). As high viral antigen load may play a role in this chronic and HBV-related carcinogenesis, the goal of HBV treatment is to eradicate viral proteins. HBV regulatory protein X (HBx) binds to the host DNA damage-binding protein 1 (DDB1) protein to degrade structural maintenance of chromosomes 5/6 (Smc5/6), resulting in activation of viral transcription from cccDNA. Here, using a split luciferase complementation assay system, we present a comprehensive compound screening system to identify inhibitors of the HBx-DDB1 interaction. Our protocol enables easy detection of interaction dynamics in real time within living cells. This technique may become a key assay to discover novel therapeutic agents for treatment of HBV infection.
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