Generation of A Stable GFP-reporter Zika Virus System for High-throughput Screening of Zika Virus Inhibitors

Zika virus(ZIKV) is associated with severe birth defects and Guillain-Barre′ syndrome and no approved vaccines or specific therapies to combat ZIKV infection are currently available. To accelerate anti-ZIKV therapeutics research, we developed a stable ZIKV GFP-reporter virus system with considerably improved GFP visibility and stability. In this system a BHK-21 cell line expressing DC-SIGNR was established to facilitate the proliferation of GFP-reporter ZIKV. Using this reporter virus system, we established a high-throughput screening assay and screened a selected plant-sourced compounds library for their ability to block ZIKV infection. More than 31 out of 974 tested compounds effectively decreased ZIKV reporter infection. Four selected compounds, homoharringtonine(HHT), bruceine D(BD), dihydroartemisinin(DHA) and digitonin(DGT), were further validated to inhibit wild-type ZIKV infection in cells of BHK-21 and human cell line A549.The FDA-approved chronic myeloid leukemia treatment drug HHT and BD were identified as broad-spectrum flavivirus inhibitors. DHA, another FDA-approved antimalarial drug effectively inhibited ZIKV infection in BHK-21 cells. HHT, BD and DHA inhibited ZIKV infection at a post-entry stage. Digitonin was found to have inhibitory activity in the early stage of viral infection. Our research provides an efficient high-throughput screening assay for ZIKV inhibitors. The active compounds identified in this study represent potential therapies for the treatment of ZIKV infection.

Control of ribosome synthesis in bacteria:the important role of rRNA chain elongation rate

Bacteria growth depends crucially on protein synthesis,which is limited by ribosome synthesis.Ribosomal RNA(rRNA)transcription is the rate-limiting step of ribosome synthesis.It is generally proposed that the transcriptional initiation rate of rRNA operon is the primary factor that controls the r RNA synthesis.In this study,we established a convenient GFP-based reporter approach for measuring the bacterial rRNA chain elongation rate.We showed that the rRNA chain elongation rate of Escherichia coli remains constant under nutrient limitation and chloramphenicol inhibition.In contrast,rRNA chain elongation rate decreases dramatically under low temperatures.Strikingly,we found that Vibrio natriegens,the fastest growing bacteria known,has a 50%higher rRNA chain elongation rate than E.coli,which contributes to its rapid ribosome synthesis.Our study demonstrates that r RNA chain elongation rate is another important factor that affects the bacterial ribosome synthesis capacity.