Alpha Enolase 1 Ubiquitination and Degradation Mediated by Ehrlichia chaffeensis TRP120 Disrupts Glycolytic Flux and Promotes Infection.
Alpha Enolase 1 Ubiquitination and Degradation Mediated by Ehrlichia chaffeensis TRP120 Disrupts Glycolytic Flux and Promotes Infection.
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沙菲埃立体TRP120介导的α烯醇化酶1泛素化和降解破坏糖酵解通量并促进感染
DOI：
10.3390/pathogens10080962
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发表时间：
2021-07-30
期刊：
影响因子：
--
通讯作者：
McBride JW
中科院分区：
文献类型：
--
作者：
Zhu B;McBride JW
查菲埃里希体调节许多宿主细胞过程，包括基因转录以促进单核吞噬细胞的感染。这些宿主细胞过程的调节是通过E. chaffeensis效应子，包括TRP 120。我们先前报道了TRP 120兼职作为HECT E3 Ub连接酶，泛素化宿主细胞转录和命运调节因子（PCGF 5和FBW 7）以促进感染。在这项研究中，我们发现了一种新的TRP 120底物，并研究了TRP 120和α-烯醇化酶（ENO 1），一种催化糖酵解途径底物脱水的金属酶之间的关系。免疫荧光显微镜和免疫共沉淀证实了TRP 120与ENO-1之间的相互作用，并在体内和体外检测到TRP 120对ENO-1的泛素化。此外，在感染过程中观察到ENO-1降解，并被蛋白酶体抑制剂硼替佐米抑制。TRP 120 HECT Ub连接酶催化位点突变体的异位表达证实了TRP 120 Ub连接酶活性在ENO-1降解中的直接作用。ENO-1的siRNA敲除与E. chaffeensis感染和ENO-1敲低通过降低丙酮酸和乳酸的水平破坏糖酵解通量，这可能有助于促进感染的宿主细胞代谢的变化。此外，我们还阐明了TRP 120自身泛素化作为一种激活事件的功能作用，它促进了UbcH 5 E2泛素结合酶的募集。这项研究进一步扩展了TRP 120底物的库，并将TRP 120 Ub连接酶在感染中的潜在作用扩展到包括代谢重编程。
Ehrlichia chaffeensis modulates numerous host cell processes, including gene transcription to promote infection of the mononuclear phagocyte. Modulation of these host cell processes is directed through E. chaffeensis effectors, including TRP120. We previously reported that TRP120 moonlights as a HECT E3 Ub ligase that ubiquitinates host cell transcription and fate regulators (PCGF5 and FBW7) to promote infection. In this study, we identified a novel TRP120 substrate and examined the relationship between TRP120 and α-enolase (ENO1), a metalloenzyme that catalyzes glycolytic pathway substrate dehydration. Immunofluorescence microscopy and coimmunoprecipitation demonstrated interaction between ENO1 and TRP120, and ubiquitination of ENO-1 by TRP120 was detected in vivo and in vitro. Further, ENO-1 degradation was observed during infection and was inhibited by the proteasomal inhibitor bortezomib. A direct role of TRP120 Ub ligase activity in ENO-1 degradation was demonstrated and confirmed by ectopic expression of TRP120 HECT Ub ligase catalytic site mutant. siRNA knockdown of ENO-1 coincided with increased E. chaffeensis infection and ENO-1 knockdown disrupted glycolytic flux by decreasing the levels of pyruvate and lactate that may contribute to changes in host cell metabolism that promote infection. In addition, we elucidated a functional role of TRP120 auto-ubiquitination as an activating event that facilitates the recruitment of the UbcH5 E2 ubiquitin-conjugating enzyme. This investigation further expands the repertoire of TRP120 substrates and extends the potential role of TRP120 Ub ligase in infection to include metabolic reprogramming.
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影响因子：
5.7
作者：
Lina TT;Farris T;Luo T;Mitra S;Zhu B;McBride JW
通讯作者：
McBride JW
影响因子：
5.7
作者：
Luo T;Dunphy PS;McBride JW
通讯作者：
McBride JW
影响因子：
3.2
作者：
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通讯作者：
Crosson, Sean
影响因子：
3.1
作者：
Dunphy, Paige Selvy;Luo, Tian;McBride, Jere W.
通讯作者：
McBride, Jere W.
影响因子：
2.9
作者：
Chen, Jo-Mei Maureen;Chiu, Shao-Chih;Yu, Chang-Tze Ricky
通讯作者：
Yu, Chang-Tze Ricky