Viral Genomics and Vaccination Unit, UMR3569 CNRS, Virology department, Institut Pasteur, 75015 Paris, France.
Viruses. 2019 Jan 15;11(1):68. doi: 10.3390/v11010068.
Flaviviruses, such as dengue (DENV), West Nile (WNV), yellow fever (YFV) and Zika (ZIKV) viruses, are mosquito-borne pathogens that present a major risk to global public health. To identify host factors that promote flavivirus replication, we performed a genome-wide gain-of-function cDNA screen for human genes that enhance the replication of flavivirus reporter particles in human cells. The screen recovered seventeen potential host proteins that promote viral replication, including the previously known dolichyl-diphosphooligosaccharide--protein glycosyltransferase non-catalytic subunit (DDOST). Using silencing approaches, we validated the role of four candidates in YFV and WNV replication: ribosomal protein L19 (RPL19), ribosomal protein S3 (RPS3), DDOST and importin 9 (IPO9). Applying a panel of virological, biochemical and microscopic methods, we validated further the role of RPL19 and DDOST as host factors required for optimal replication of YFV, WNV and ZIKV. The genome-wide gain-of-function screen is thus a valid approach to advance our understanding of flavivirus replication.
黄病毒，如登革热（DENV）、西尼罗河（WNV）、黄热病（YFV）和寨卡（ZIKV）病毒，是通过蚊子传播的病原体，对全球公共卫生构成重大威胁。为了鉴定促进黄病毒复制的宿主因素，我们进行了全基因组功能获得 cDNA 筛选，以鉴定在人类细胞中增强黄病毒报告颗粒复制的人类基因。该筛选回收了十七种可能促进病毒复制的潜在宿主蛋白，包括先前已知的双萜醇磷酸寡糖-蛋白糖基转移酶非催化亚基（DDOST）。通过沉默方法，我们验证了四个候选基因在 YFV 和 WNV 复制中的作用：核糖体蛋白 L19（RPL19）、核糖体蛋白 S3（RPS3）、DDOST 和导入蛋白 9（IPO9）。应用一系列病毒学、生化和显微镜方法，我们进一步验证了 RPL19 和 DDOST 作为宿主因子在 YFV、WNV 和 ZIKV 最佳复制中的作用。因此，全基因组功能获得筛选是一种有效的方法，可以深入了解黄病毒的复制。
