College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Key Laboratory of Silkworm and Bee Resource Utilization and Innovation of Zhejiang Province, Hangzhou 310000, China.
Viruses. 2022 Jan 14;14(1):153. doi: 10.3390/v14010153.
Hyperexpression of and , two very late genes, is one of the remarkable characteristics in the baculovirus life cycle. However, the mechanisms underlying the hyperexpression of these two genes are still incompletely understood. In this study, actin was identified as a highly potential binding partner of and promoters by conducting DNA pull-down and LC-MS/MS analyses. Inhibiting actin dynamics delayed and decreased the transcription of and . Actin interacted with viral RNA polymerase and transcription regulators, and the nuclear import of viral polymerase was inhibited with the disruption of actin dynamics. Simultaneously, the high enrichment of actin in and promoters discovered via a chromatin immunoprecipitation (ChIP) assay indicated that actin was a component of the viral polymerase TIC. Moreover, overexpression of actin surprisingly upregulated the expression of luciferase (Luc) under the control of and promoters. Taken together, actin participated in the hyperexpression of and as a component of TIC. These results facilitate the promotion of the expression efficiency of foreign genes in the baculovirus expression vector system (BEVS).
过度表达和 ,这两个非常晚期的基因,是杆状病毒生命周期的显著特征之一。然而,这两个基因过度表达的机制仍不完全清楚。在这项研究中,通过 DNA 下拉和 LC-MS/MS 分析,肌动蛋白被鉴定为和启动子的一个高度潜在的结合伙伴。抑制肌动蛋白动力学延迟和降低了和的转录。肌动蛋白与病毒 RNA 聚合酶和转录调节剂相互作用,并且肌动蛋白动力学的破坏抑制了病毒聚合酶的核输入。同时,通过染色质免疫沉淀 (ChIP) 实验发现肌动蛋白在和启动子中高度富集,表明肌动蛋白是病毒聚合酶 TIC 的组成部分。此外,肌动蛋白的过表达出人意料地上调了在和启动子控制下的荧光素酶 (Luc) 的表达。综上所述,肌动蛋白作为 TIC 的一部分参与了和的过度表达。这些结果有助于提高杆状病毒表达载体系统 (BEVS) 中外源基因的表达效率。