Acta Virol. 2023;67(1):42-50. doi: 10.4149/av_2023_104.
Late expression factor 11 (LEF-11) is an essential protein in the regulation of Bombyx mori nucleopolyhedrovirus (BmNPV) DNA replication and late gene expression. Our recent quantitative analysis of protein acetylome revealed for the first time that LEF-11 can be acetylated at one lysine residue (K83) during viral infection, but the underlying mechanism is unclear. The acetylation level for K83 was down-regulated after 36 h post-infection by approximately 30%. To clarify the regulatory function of this modification, overlap PCR was used for site-specific mutagenesis for acetylated (K83Q) or deacetylated (K83R) mimic mutants of LEF-11. The results of viral titration and quantitative polymerase chain reaction showed that after K83 acetylation, budding virion production and the viral genome replication level were significantly upregulated. Meanwhile, the results of yeast two-hybrid (Y2H) system confirmed that K83 deacetylation modification inhibited the interaction between LEF-11 and immediate early gene 1 (IE-1). In conclusion, the acetylation of LEF-11 at K83 might enhance the interaction with IE-1 in the host cell nucleus to promote viral DNA replication, and might be one of the antiviral strategies of the silkworm host. The host inhibits virus proliferation by deacetylating LEF-11. Keywords: BmNPV; LEF-11; acetylation; virus replication; protein interaction.
晚期表达因子 11(LEF-11)是调控家蚕核型多角体病毒(BmNPV)DNA 复制和晚期基因表达的必需蛋白。我们最近对蛋白质乙酰化组的定量分析首次表明,LEF-11 在病毒感染过程中可以在一个赖氨酸残基(K83)上发生乙酰化,但具体的调控机制尚不清楚。感染后 36 小时,K83 的乙酰化水平下降了约 30%。为了阐明这种修饰的调控功能,采用重叠 PCR 对 LEF-11 的乙酰化(K83Q)或去乙酰化(K83R)模拟突变体进行了定点突变。病毒滴定和定量聚合酶链反应的结果表明,K83 乙酰化后,出芽病毒粒子的产生和病毒基因组的复制水平显著上调。同时,酵母双杂交(Y2H)系统的结果证实,K83 去乙酰化修饰抑制了 LEF-11 与早期基因 1(IE-1)之间的相互作用。综上所述,LEF-11 在 K83 上的乙酰化可能增强了其与宿主细胞核中 IE-1 的相互作用,从而促进病毒 DNA 的复制,这可能是宿主抗病毒的策略之一。宿主通过去乙酰化 LEF-11 抑制病毒的增殖。关键词:BmNPV;LEF-11;乙酰化;病毒复制;蛋白质相互作用。