Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai, People's Republic of China.
PLoS One. 2013 Jul 24;8(7):e69442. doi: 10.1371/journal.pone.0069442. Print 2013.
Histone H3-lysine(9) (H3K9) trimethyltransferase gene Su(var) 3-9 was cloned and identified in three Spodoptera insects, Spodopterafrugiperda (S. frugiperda), S. exigua and S. litura. Sequence analysis showed that Spodoptera Su(var) 3-9 is highly conserved evolutionarily. Su(var) 3-9 protein was found to be localized in the nucleus in Sf9 cells, and interact with histone H3, and the heterochromatin protein 1a (HP1a) and HP1b. A dose-dependent enzymatic activity was found at both 27 °C and 37 °C in vitro, with higher activity at 27 °C. Addition of specific inhibitor chaetocin resulted in decreased histone methylation level and host chromatin relaxation. In contrast, overexpression of Su(var) 3-9 caused increased histone methylation level and cellular genome compaction. In AcMNV-infected Sf9 cells, the transcription of Su(var) 3-9 increased at late time of infection, although the mRNA levels of most cellular genes decreased. Pre-treatment of Sf9 cells with chaetocin speeded up viral DNA replication, and increased the transcription level of a variety of virus genes, whereas in Sf9 cells pre-transformed with Su(var) 3-9 expression vector, viral DNA replication slow down slightly. These findings suggest that Su(var) 3-9 might participate in the viral genes expression an genome replication repression during AcMNPV infection. It provided a new insight for the understanding virus-host interaction mechanism.
组蛋白 H3-赖氨酸(9)(H3K9)三甲基转移酶基因 Su(var)3-9 在三种夜蛾昆虫 Spodoptera frugiperda(S. frugiperda)、S. exigua 和 S. litura 中被克隆和鉴定。序列分析表明,Spodoptera Su(var)3-9 在进化上高度保守。在 Sf9 细胞中发现 Su(var)3-9 蛋白定位于细胞核中,并与组蛋白 H3、异染色质蛋白 1a(HP1a)和 HP1b 相互作用。在体外,在 27°C 和 37°C 下均发现具有剂量依赖性的酶活性,在 27°C 时活性更高。添加特异性抑制剂 chaetocin 导致组蛋白甲基化水平降低和宿主染色质松弛。相比之下,Su(var)3-9 的过表达导致组蛋白甲基化水平增加和细胞基因组紧缩。在 AcMNV 感染的 Sf9 细胞中,Su(var)3-9 的转录在感染后期增加,尽管大多数细胞基因的 mRNA 水平下降。在 Sf9 细胞中用 chaetocin 预处理可加速病毒 DNA 复制,并增加多种病毒基因的转录水平,而在预先转化了 Su(var)3-9 表达载体的 Sf9 细胞中,病毒 DNA 复制略有减缓。这些发现表明,Su(var)3-9 可能参与 AcMNPV 感染过程中的病毒基因表达和基因组复制抑制。它为理解病毒-宿主相互作用机制提供了新的视角。