Forward genetics in Wolbachia: Regulation of Wolbachia proliferation by the amplification and deletion of an addictive genomic island.
Forward genetics in Wolbachia: Regulation of Wolbachia proliferation by the amplification and deletion of an addictive genomic island.
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沃尔巴克氏菌的正向遗传学：通过扩增和删除成瘾基因组岛来调节沃尔巴克氏菌的增殖。
DOI：
10.1371/journal.pgen.1009612
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发表时间：
2021-06
期刊：
影响因子：
4.5
通讯作者：
Teixeira L
中科院分区：
文献类型：
--
作者：
Duarte EH;Carvalho A;López-Madrigal S;Costa J;Teixeira L
沃尔巴克氏体是最普遍的细菌内共生体之一，感染约40%的陆生节肢动物物种。沃尔巴克氏体通常是一种生殖寄生虫，但也可以为其宿主提供健康益处，例如，保护免受病毒病原体的侵害。这种保护作用目前正通过释放感染沃尔巴克氏体的蚊子来对抗虫媒病毒的传播。滴度调节是沃尔巴克氏体生物学的一个重要方面。更高的滴度可以导致更强的表型和传播的保真度，但对宿主来说可能具有更高的成本。由于沃尔巴克氏体是母系传播的，它的适应性取决于宿主的适应性，因此，它对宿主的消耗可能是选择的结果。了解沃尔巴克氏体滴度是如何调节的以及沃尔巴克氏体生物学的其他方面一直受到缺乏遗传工具的阻碍。在这里，我们开发了一个正向遗传筛选，以确定新的沃尔巴克氏体过度增殖突变体。我们详细描述了两个新的突变体，wMelPop2和wMelOctoless，并表明Octomom基因组区域的扩增或丢失导致过度增殖。这些结果证实了以前的数据，并扩大了这个基因组区域在控制沃尔巴克氏体增殖的复杂作用。这两种新的突变体都缩短了宿主的寿命，并增加了抗病毒保护。此外，我们表明，沃尔巴克氏体在果蝇的增殖率取决于Octomom拷贝数，宿主发育阶段和温度之间的相互作用。我们的分析还表明，沃尔巴克氏体的寿命缩短和抗病毒保护表型是依赖于不同的，但相关的，属性的内共生体的增殖率和滴度感染时，分别。我们还证明了一种新的和公正的实验方法来研究沃尔巴克氏体生物学的可行性，这可以进一步适应其他遗传上难治性细菌内共生体的特点。昆虫通常携带生活在其细胞内的细菌，并从母体传播给后代。沃尔巴克氏体是最常见的细菌之一，可以强烈影响昆虫生物学。它保护一些宿主免受病毒感染的能力正在通过将沃尔巴克氏体引入这些昆虫来对抗蚊子传播的病毒。宿主体内沃尔巴克氏体的数量会影响它们的相互作用，必须加以很好的控制。为了理解这一过程，我们筛选了沃尔巴克氏体的新突变体，这些突变体在果蝇中过度增殖。我们确定了两个突变体，并详细描述了它们的特征。一个突变体有太多的一组特定基因的拷贝，证实了以前的类似结果。然而，另一个突变的沃尔巴克氏体失去了这些完全相同的基因，表明它们在生长调节中特别重要。此外，我们表明，不同的沃尔巴克氏体变体的增殖取决于温度，和昆虫宿主的发育阶段。最后，数据表明，保护病毒和成本的沃尔巴克氏体依赖于相关的，但不同的方面，这种控制的增长。总之，我们表明，我们可以筛选新的突变体的沃尔巴克氏体和更好地了解如何控制生长的遗传控制沃尔巴克氏体。
Wolbachia is one of the most prevalent bacterial endosymbionts, infecting approximately 40% of terrestrial arthropod species. Wolbachia is often a reproductive parasite but can also provide fitness benefits to its host, as, for example, protection against viral pathogens. This protective effect is currently being applied to fight arboviruses transmission by releasing Wolbachia-transinfected mosquitoes. Titre regulation is a crucial aspect of Wolbachia biology. Higher titres can lead to stronger phenotypes and fidelity of transmission but can have a higher cost to the host. Since Wolbachia is maternally transmitted, its fitness depends on host fitness, and, therefore, its cost to the host may be under selection. Understanding how Wolbachia titres are regulated and other aspects of Wolbachia biology has been hampered by the lack of genetic tools. Here we developed a forward genetic screen to identify new Wolbachia over-proliferative mutant variants. We characterized in detail two new mutants, wMelPop2 and wMelOctoless, and show that the amplification or loss of the Octomom genomic region lead to over-proliferation. These results confirm previous data and expand on the complex role of this genomic region in the control of Wolbachia proliferation. Both new mutants shorten the host lifespan and increase antiviral protection. Moreover, we show that Wolbachia proliferation rate in Drosophila melanogaster depends on the interaction between Octomom copy number, the host developmental stage, and temperature. Our analysis also suggests that the life shortening and antiviral protection phenotypes of Wolbachia are dependent on different, but related, properties of the endosymbiont; the rate of proliferation and the titres near the time of infection, respectively. We also demonstrate the feasibility of a novel and unbiased experimental approach to study Wolbachia biology, which could be further adapted to characterize other genetically intractable bacterial endosymbionts. Insects often carry bacteria that live within their cells and are transmitted from the mother to the progeny. Wolbachia is one the most common of such bacteria and can strongly influence the insect biology. Its capacity to protect some hosts from viral infection is being used in the fight against mosquitoes-transmitted viruses by introducing Wolbachia in these insects. The amount of Wolbachia within the host can impact their interaction and must be well controlled. To understand this process we screened for new mutants of Wolbachia that proliferate too much in the fruit fly Drosophila melanogaster. We identified two mutants and characterized them in detail. One mutant has too many copies of a specific set of genes, confirming previous similar results. However, the other mutant Wolbachia lost those exact same genes, showing that they are particularly important in growth regulation. Moreover, we show that proliferation of different Wolbachia variants depends on temperature, and the developmental stage of the insect host. Finally, the data indicate that protection to viruses and cost of Wolbachia depend on related but different aspects of this control of growth. In summary, we show that we can screen for new mutants of Wolbachia and understand better how control of growth is genetically controlled by Wolbachia.
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