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鉴定和表征夜蛾 Su(var)3-9 组蛋白 H3K9 三甲基转移酶及其在 AcMNPV 感染中的作用。

Identification and characterization of the Spodoptera Su(var) 3-9 histone H3K9 trimethyltransferase and its effect in AcMNPV infection.

机构信息

Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai, People's Republic of China.

出版信息

PLoS One. 2013 Jul 24;8(7):e69442. doi: 10.1371/journal.pone.0069442. Print 2013.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3794/3722159/e36244e4cfc7/pone.0069442.g001.jpg

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