Computational prediction and experimental validation of Salmonella Typhimurium SopE-mediated fine-tuning of autophagy in intestinal epithelial cells.
Computational prediction and experimental validation of Salmonella Typhimurium SopE-mediated fine-tuning of autophagy in intestinal epithelial cells.
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DOI：
10.3389/fcimb.2022.834895
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发表时间：
2022
影响因子：
5.7
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中科院分区：
文献类型：
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大自噬是真核细胞中普遍存在的稳态和促进健康的再循环过程，靶向错误折叠的蛋白质、受损的细胞器和细胞内感染因子。一些细胞内病原体，如沙门氏菌血清型鼠伤寒在发病过程中劫持这一过程。在这里，我们研究宿主转录因子和沙门氏菌分泌的效应蛋白之间的潜在蛋白质-蛋白质相互作用及其对宿主基因转录的影响。一项系统水平的分析确定了有可能影响核心自噬基因调节的沙门氏菌效应蛋白。SPI-1效应蛋白，SopE，这是预测与自噬过程的调节蛋白相互作用的影响，进行了研究，以验证我们的方法。然后，我们通过实验证实，SopE可以直接结合SP1，这是一种宿主转录因子，它调节自噬基因MAP 1 LC 3B的表达。我们还发现，SopE可能在调节自噬中具有双重作用：在沙门氏菌感染引发的MAP 1 LC 3B转录的初始增加之后，随后在感染后6小时MAP 1 LC 3B转录的降低是SopE依赖的。SopE还在调节自噬通量机制中发挥作用，特别是MAP 1 LC 3B和p62自噬蛋白，这取决于已经发生的自噬水平。在典型的上皮细胞感染后，自噬通量增加。然而，当自噬在感染前被化学诱导时，SopE抑制了自噬通量。当大多数细胞内沙门氏菌细胞与SCV（缺乏sifA的菌株）不相关时，也观察到相同的情况，而不管感染前的自噬诱导状态如何。我们展示了如何使用调控网络分析来更好地分析致病性效应蛋白的影响，在这种情况下，沙门氏菌。这项研究补充了以前的工作，我们已经证明，特定的病原体效应物可以通过与自噬蛋白直接相互作用来影响自噬过程。在这里，我们表明，效应蛋白也可以影响上游调控的过程。这种跨学科的研究可以增加我们对感染过程的理解，并指出肠上皮细胞防御中重要的靶点。
Macroautophagy is a ubiquitous homeostasis and health-promoting recycling process of eukaryotic cells, targeting misfolded proteins, damaged organelles and intracellular infectious agents. Some intracellular pathogens such as Salmonella enterica serovar Typhimurium hijack this process during pathogenesis. Here we investigate potential protein-protein interactions between host transcription factors and secreted effector proteins of Salmonella and their effect on host gene transcription. A systems-level analysis identified Salmonella effector proteins that had the potential to affect core autophagy gene regulation. The effect of a SPI-1 effector protein, SopE, that was predicted to interact with regulatory proteins of the autophagy process, was investigated to validate our approach. We then confirmed experimentally that SopE can directly bind to SP1, a host transcription factor, which modulates the expression of the autophagy gene MAP1LC3B. We also revealed that SopE might have a double role in the modulation of autophagy: Following initial increase of MAP1LC3B transcription triggered by Salmonella infection, subsequent decrease in MAP1LC3B transcription at 6h post-infection was SopE-dependent. SopE also played a role in modulation of the autophagy flux machinery, in particular MAP1LC3B and p62 autophagy proteins, depending on the level of autophagy already taking place. Upon typical infection of epithelial cells, the autophagic flux is increased. However, when autophagy was chemically induced prior to infection, SopE dampened the autophagic flux. The same was also observed when most of the intracellular Salmonella cells were not associated with the SCV (strain lacking sifA) regardless of the autophagy induction status before infection. We demonstrated how regulatory network analysis can be used to better characterise the impact of pathogenic effector proteins, in this case, Salmonella. This study complements previous work in which we had demonstrated that specific pathogen effectors can affect the autophagy process through direct interaction with autophagy proteins. Here we show that effector proteins can also influence the upstream regulation of the process. Such interdisciplinary studies can increase our understanding of the infection process and point out targets important in intestinal epithelial cell defense.
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影响因子：
4.6
作者：
Han H;Shim H;Shin D;Shim JE;Ko Y;Shin J;Kim H;Cho A;Kim E;Lee T;Kim H;Kim K;Yang S;Bae D;Yun A;Kim S;Kim CY;Cho HJ;Kang B;Shin S;Lee I
通讯作者：
Lee I
影响因子：
3.4
作者：
Guidi, Riccardo;Levi, Laura;Frisan, Teresa
通讯作者：
Frisan, Teresa
影响因子：
4.4
作者：
Bovolenta LA;Acencio ML;Lemke N
通讯作者：
Lemke N
影响因子：
4.1
作者：
Herhaus, Lina;Dikic, Ivan
通讯作者：
Dikic, Ivan
影响因子：
3.1
作者：
Brumell, JH;Tang, P;Finlay, BB
通讯作者：
Finlay, BB