RNAi screen in Drosophila cells reveals the involvement of the Tom complex in Chlamydia infection.
RNAi screen in Drosophila cells reveals the involvement of the Tom complex in Chlamydia infection.
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DOI：
10.1371/journal.ppat.0030155
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发表时间：
2007-10-26
期刊：
影响因子：
6.7
通讯作者：
Agaisse H
中科院分区：
文献类型：
--
作者：
Derré I;Pypaert M;Dautry-Varsat A;Agaisse H
衣原体属物种是细胞内专性细菌病原体，感染广泛的宿主细胞。在这里，我们表明，C。在果蝇SL 2细胞中，细胞进入、复制并执行一个完整的发育周期。使用这个模型系统，我们进行了全基因组RNA干扰筛选，并确定了54个因子，当耗尽时，抑制C。败血症感染通过测试每个候选人的击倒对L。单核细胞增多症感染，我们已经确定了31个候选人可能是C。败血症感染我们发现的因素预期有影响衣原体感染，如硫酸乙酰肝素糖胺聚糖和肌动蛋白和微管重塑因子。我们还确定了以前没有描述为参与衣原体感染的因素。例如，我们鉴定了Tim-Tom复合体的成员，这是一种多蛋白复合体，参与识别和将核编码蛋白输入线粒体，这是C。果蝇细胞的感染。最后，我们证实了Tom 40或Tom 22的缺失也降低了C。在哺乳动物细胞中的细菌感染。然而，C.沙眼衣原体感染不受影响，提示其机制是C.特定于鳕鱼。 衣原体属物种是细胞内细菌病原体，感染广泛的宿主并引起各种疾病，包括发展中国家可预防的失明、性传播疾病和肺炎。衣原体属物种能够在宿主细胞内建立其复制小生境，驻留在膜结合区室中，该膜结合区室用作抵抗免疫监视和抗微生物剂的保护屏障，但也用作与宿主细胞交换因子的“过滤器”。尽管衣原体对人类健康至关重要，但人们对感染过程的机制知之甚少。衣原体致病机制的研究是具有挑战性的，因为衣原体属。不适于遗传操作，并且难以在哺乳动物宿主中进行广泛的遗传方法。为了克服这些困难，我们使用果蝇细胞来模拟衣原体感染。我们进行了全基因组RNA干扰筛选，并确定了宿主因子，当耗尽，减少衣原体感染。为了验证我们的方法，我们进一步证明了所鉴定的因子也是感染哺乳动物细胞所必需的。这项工作将帮助我们更好地了解衣原体与其宿主之间的复杂相互作用，并可能确定新的治疗靶点。
Chlamydia spp. are intracellular obligate bacterial pathogens that infect a wide range of host cells. Here, we show that C. caviae enters, replicates, and performs a complete developmental cycle in Drosophila SL2 cells. Using this model system, we have performed a genome-wide RNA interference screen and identified 54 factors that, when depleted, inhibit C. caviae infection. By testing the effect of each candidate's knock down on L. monocytogenes infection, we have identified 31 candidates presumably specific of C. caviae infection. We found factors expected to have an effect on Chlamydia infection, such as heparansulfate glycosaminoglycans and actin and microtubule remodeling factors. We also identified factors that were not previously described as involved in Chlamydia infection. For instance, we identified members of the Tim-Tom complex, a multiprotein complex involved in the recognition and import of nuclear-encoded proteins to the mitochondria, as required for C. caviae infection of Drosophila cells. Finally, we confirmed that depletion of either Tom40 or Tom22 also reduced C. caviae infection in mammalian cells. However, C. trachomatis infection was not affected, suggesting that the mechanism involved is C. caviae specific. Chlamydia spp. are intracellular bacterial pathogens that infect a wide range of hosts and cause various diseases, including preventable blindness in developing countries, sexually transmitted disease, and pneumonia. Chlamydia spp. are able to establish their replication niche inside the host cell, residing in a membrane-bound compartment that serves as a protector shield against immune surveillance and antimicrobial agents but also acts as a “filter” to exchange factors with the host cell. Despite the primary importance of Chlamydia for human health, little is known about the mechanisms underlying the infection process. The study of Chlamydia pathogenesis is challenging because Chlamydia spp. are not amenable to genetic manipulation and it is difficult to conduct extensive genetic approaches in the mammalian host. To circumvent these difficulties, we have used Drosophila cells to model Chlamydia infection. We conducted a genome-wide RNA interference screen and identified host factors that, when depleted, reduce Chlamydia infection. Validating our approach, we further showed that the identified factors were also required for infection in mammalian cells. This work will help us better understand the complex interaction between Chlamydia and its host and potentially identify novel targets for therapeutic treatment.
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影响因子：
3.4
作者：
Cheng, LW;Portnoy, DA
通讯作者：
Portnoy, DA
DOI：
10.1073/pnas.0506461102
发表时间：
2005-09-20
影响因子：
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DOI：
10.1073/pnas.110149597
发表时间：
2000-06-06
影响因子：
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作者：
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通讯作者：
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作者：
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