Institute of Virology, University of Zurich, Zurich, Switzerland.
Department for Chemistry, Biochemistry and Pharmaceutical Sciences, University of Bern, Bern, Switzerland.
PLoS Pathog. 2021 Jun 1;17(6):e1009638. doi: 10.1371/journal.ppat.1009638. eCollection 2021 Jun.
Adeno-associated virus (AAV) genome replication only occurs in the presence of a co-infecting helper virus such as adenovirus type 5 (AdV5) or herpes simplex virus type 1 (HSV-1). AdV5-supported replication of the AAV genome has been described to occur in a strand-displacement rolling hairpin replication (RHR) mechanism initiated at the AAV 3' inverted terminal repeat (ITR) end. It has been assumed that the same mechanism applies to HSV-1-supported AAV genome replication. Using Southern analysis and nanopore sequencing as a novel, high-throughput approach to study viral genome replication we demonstrate the formation of double-stranded head-to-tail concatemers of AAV genomes in the presence of HSV-1, thus providing evidence for an unequivocal rolling circle replication (RCR) mechanism. This stands in contrast to the textbook model of AAV genome replication when HSV-1 is the helper virus.
腺相关病毒 (AAV) 基因组复制仅在辅助病毒（如腺病毒 5 型 (AdV5) 或单纯疱疹病毒 1 型 (HSV-1)）的共感染下发生。腺病毒 5 型支持的 AAV 基因组复制已被描述为在 AAV 3' 反向末端重复序列 (ITR) 末端起始的链置换滚动发夹复制 (RHR) 机制中发生。人们假设相同的机制适用于 HSV-1 支持的 AAV 基因组复制。使用 Southern 分析和纳米孔测序作为一种新颖的高通量方法来研究病毒基因组复制，我们证明了在 HSV-1 存在的情况下 AAV 基因组双链头尾连接的形成，从而为明确的滚环复制 (RCR) 机制提供了证据。这与 HSV-1 作为辅助病毒时 AAV 基因组复制的教科书模型形成对比。
