Characterization of Episomal Replication of Bovine Papillomavirus Type 1 DNA in Long-Term Virion-Infected Saccharomyces Cerevisiae Culture  

作　　者：Quanmei Tu Weixu Feng Zhuo Chen Qijia Li Yu Zhao Jun Chen Pengfei Jiang Xiangyang Xue Lifang Zhang Kong-Nan Zhao 

机构地区：[1]Department of Obstetrics and Gynaecology,The Second Affiliated Hospital and Yuyin Children Hospital of Wenzhou Medical University,Wenzhou,325035,China [2]Institute of Molecular Virology and Immunology,Department of Microbiology and Immunology,School of Basic Medical Sciences,Wenzhou Medical University,Wenzhou,325035,China [3]Australian Institute for Bioengineering and Nanotechnology,the University of Queensland,St Lucia,4067,Australia

出　　处：《Virologica Sinica》2021年第6期1492-1502,共11页中国病毒学（英文版）

基　　金：This work was funded in part by grants from the National Nature Science Foundation of China(81772791 and 81172463)。

摘　　要：We have previously reported that bovine papillomavirus type 1(BPV-1) DNA can replicate its genome and produce infectious virus-like particles in short term virion-infected S. cerevisiae(budding yeast) cultures(Zhao and Frazer 2002,Journal of Virology, 76:3359–64 and 76:12265–73). Here, we report the episomal replications of BPV-1 DNA in long term virion-infected S. cerevisiae culture up to 108 days. Episomal replications of the BPV-1 DNA could be divided into three patterns at three stages, early active replication(day 3–16), middle weak replication(day 23–34/45) and late stable replication(day 45–82). Two-dimensional gel electrophoresis analysis and Southern blot hybridization have revealed further that multiple replication intermediates of BPV-1 DNA including linear form, stranded DNA, monomers and higher oligomers were detected in the virion-infected yeast cells over the time course. Higher oligomers shown as covalently closed circular DNAs(cccDNAs) are the most important replication intermediates that serve as the main nuclear transcription template for producing all viral RNAs in the viral life cycle. In this study, the cccDNAs were generated at the early active replication stage with the highest frequencies and then at late stable replication, but they appeared to be suppressed at the middle weak replication. Our data provided a novel insight that BPV-1 genomic DNA could replicate episomally for the long period and produce the key replication intermediates cccDNAs in S. cerevisiae system.

关 键 词：Bovine papillomavirus type 1(BPV-1) Saccharomyces cerevisiae Genomic DNA replication Agarose gel electrophoresis Southern blot hybridization 

分 类 号：Q939.4[生物学—微生物学]