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丽蝇蛹集金小蜂毒液腺转录组凸显了新基因在毒液功能中的重要性。

The venom gland transcriptome of the parasitoid wasp Nasonia vitripennis highlights the importance of novel genes in venom function.

作者信息

Sim Andre D, Wheeler David

机构信息

Institute of Fundamental Sciences, Massey University, Palmerston North, 4442, New Zealand.

出版信息

BMC Genomics. 2016 Aug 8;17:571. doi: 10.1186/s12864-016-2924-7.

Abstract

BACKGROUND

Prior to egg laying the parasitoid wasp Nasonia vitripennis envenomates its pupal host with a complex mixture of venom peptides. This venom induces several dramatic changes in the host, including developmental arrest, immunosuppression, and altered metabolism. The diverse and potent bioactivity of N. vitripennis venom provides opportunities for the development of novel acting pharmaceuticals based on these molecules. However, currently very little is known about the specific functions of individual venom peptides or what mechanisms underlie the hosts response to envenomation. Many of the venom peptides also lack bioinformatically derived annotations because no homologs can be identified in the sequences databases. The RNA interference system of N. vitripennis provides a method for functional characterisation of venom protein encoding genes, however working with the current list of 79 candidates represents a daunting task. For this reason we were interested in determining the expression levels of venom encoding genes in the venom gland, as this information could be used to rank candidates for further study. To do this we carried out deep transcriptome sequencing of the venom gland and ovary tissue and used RNA-seq to rank the venom protein encoding genes by expression level. The generation of a specific venom gland transcriptome dataset also provides further opportunities to investigate novel features of this specialised organ.

RESULTS

RNA-seq revealed that the highest expressed venom encoding gene in the venom gland was 'Venom protein Y'. The highest expressed annotated gene in this tissue was serine protease Nasvi2EG007167, which has previously been implicated in the apoptotic activity of N. vitripennis venom. As expected the RNA-seq confirmed that venom encoding genes are almost exclusively expressed in the venom gland relative to the neighbouring ovary tissue. Novel genes appear to perform key roles in N. vitripennis venom function, with over half of the 15 highest expressed venom encoding loci lacking bioinformatic annotations. The high throughput sequencing data also provided evidence for the existence of an additional 472 previously undescribed transcribed regions in the N. vitripennis genome. Finally, metatranscriptomic analysis of the venom gland transcriptome finds little evidence for the role of Wolbachia in the venom system.

CONCLUSIONS

The expression level information provided here for the N. vitripennis venom protein encoding genes represents a valuable dataset that can be used by the research community to rank candidates for further functional characterisation. These candidates represent bioactive peptides valuable in the development of new pharmaceuticals.

摘要

背景

在产卵前,丽蝇蛹集金小蜂会用一种复杂的毒液肽混合物对其蛹宿主进行毒液注射。这种毒液会在宿主体内引发多种显著变化,包括发育停滞、免疫抑制和代谢改变。丽蝇蛹集金小蜂毒液多样且强大的生物活性为基于这些分子开发新型药物提供了机会。然而,目前对于单个毒液肽的具体功能或宿主对毒液注射反应的潜在机制知之甚少。许多毒液肽在生物信息学上也缺乏注释,因为在序列数据库中无法识别其同源物。丽蝇蛹集金小蜂的RNA干扰系统为毒液蛋白编码基因的功能表征提供了一种方法,然而处理当前的79个候选基因列表是一项艰巨的任务。因此,我们有兴趣确定毒液腺中毒液编码基因的表达水平,因为这些信息可用于对候选基因进行排名以便进一步研究。为此,我们对毒液腺和卵巢组织进行了深度转录组测序,并使用RNA测序按表达水平对毒液蛋白编码基因进行排名。特定毒液腺转录组数据集的生成也为研究这个特殊器官的新特征提供了更多机会。

结果

RNA测序显示,毒液腺中表达最高的毒液编码基因是“毒液蛋白Y”。该组织中表达最高的注释基因是丝氨酸蛋白酶Nasvi2EG007167,此前已发现它与丽蝇蛹集金小蜂毒液的凋亡活性有关。正如预期的那样,RNA测序证实毒液编码基因相对于相邻的卵巢组织几乎只在毒液腺中表达。新基因似乎在丽蝇蛹集金小蜂毒液功能中发挥关键作用,在表达最高的15个毒液编码基因座中,超过一半缺乏生物信息学注释。高通量测序数据还为丽蝇蛹集金小蜂基因组中另外472个先前未描述的转录区域的存在提供了证据。最后,对毒液腺转录组的宏转录组分析几乎没有发现沃尔巴克氏体在毒液系统中起作用的证据。

结论

这里提供的丽蝇蛹集金小蜂毒液蛋白编码基因的表达水平信息是一个有价值的数据集,研究界可利用它对候选基因进行排名以便进一步进行功能表征。这些候选基因代表了对开发新药物有价值的生物活性肽。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e1a3/4977848/9c1bf09ddf5b/12864_2016_2924_Fig1_HTML.jpg

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