Kumar Sunil, Jung Jin-Kyo, Kim Yonggyun
Department of Plant Medicals, Andong National University, Andong, Korea.
Division of Crop Cultivation and Environment Research, Department of Central Area Crop Science, National Institute of Crop Science, Rural Development Administration, Suwon, Korea.
PLoS One. 2017 May 9;12(5):e0177066. doi: 10.1371/journal.pone.0177066. eCollection 2017.
A viral histone H4 (CpBV-H4) is encoded in a polydnavirus, Cotesia plutellae bracovirus (CpBV). It plays a crucial role in parasitism of an endoparasitoid wasp, C. plutellae, against diamondback moth, Plutella xylostella, by altering host gene expression in an epigenetic mode by its N-terminal tail after joining host nucleosomes. Comparative transcriptomic analysis between parasitized and nonparasitized P. xylostella by RNA-Seq indicated that 1,858 genes were altered at more than two folds in expression levels at late parasitic stage, including 877 up-regulated genes and 981 down-regulated genes. Among parasitic factors altering host gene expression, CpBV-H4 alone explained 16.3% of these expressional changes. To characterize the joining sites of CpBV-H4 on host chromosomes, ChIP-Seq (chromatin immunoprecipitation followed by deep sequencing) was applied to chromatins extracted from parasitized larvae. It identified specific 538 ChIP targets. Joining sites were rich (60.2%) in AT sequence. Almost 40% of ChIP targets included short nucleotide repeat sequences presumably recognizable by transcriptional factors and chromatin remodeling factors. To further validate these CpBV-H4 targets, CpBV-H4 was transiently expressed in nonparasitized host at late larval stage and subjected to ChIP-Seq. Two kinds of ChIP-Seqs shared 51 core joining sites. Common targets were close (within 1 kb) to genes regulated at expression levels by CpBV-H4. However, other host genes not close to CpBV-H4 joining sites were also regulated by CpBV-H4. These results indicate that CpBV-H4 joins specific chromatin regions of P. xylostella and controls about one sixth of the total host genes that were regulated by C. plutellae parasitism in an epigenetic mode.
一种病毒组蛋白H4(小菜蛾茧蜂杆状病毒组蛋白H4,CpBV-H4)编码于一种多DNA病毒——小菜蛾茧蜂杆状病毒(CpBV)中。在一种内寄生蜂——小菜蛾茧蜂寄生小菜蛾的过程中,它通过在加入宿主核小体后利用其N端尾巴以表观遗传模式改变宿主基因表达,发挥着关键作用。通过RNA测序对被寄生和未被寄生的小菜蛾进行比较转录组分析表明,在寄生后期,1858个基因的表达水平变化超过两倍,其中包括877个上调基因和981个下调基因。在改变宿主基因表达的寄生因子中,仅CpBV-H4就解释了这些表达变化的16.3%。为了确定CpBV-H4在宿主染色体上的结合位点,染色质免疫沉淀测序(ChIP-Seq)被应用于从被寄生幼虫中提取的染色质。它确定了538个特定的ChIP靶点。结合位点富含AT序列(占60.2%)。几乎40%的ChIP靶点包含可能被转录因子和染色质重塑因子识别的短核苷酸重复序列。为了进一步验证这些CpBV-H4靶点,在幼虫后期将CpBV-H4在未被寄生的宿主中瞬时表达,并进行ChIP-Seq。两种ChIP-Seq共有51个核心结合位点。共同靶点与在表达水平上受CpBV-H4调控的基因距离较近(在1 kb内)。然而,其他与CpBV-H4结合位点距离不近的宿主基因也受CpBV-H4调控。这些结果表明,CpBV-H4与小菜蛾的特定染色质区域结合,并以表观遗传模式控制着受小菜蛾寄生调控的约六分之一的宿主基因总数。