Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, OH, USA.
Mol Microbiol. 2021 Feb;115(2):305-319. doi: 10.1111/mmi.14617. Epub 2020 Nov 5.
Many Gram-negative pathogens use a type III secretion system (T3SS) to promote disease by injecting effector proteins into host cells. Common to many T3SSs is that injection of effector proteins is feedback inhibited. The mechanism of feedback inhibition and its role in pathogenesis are unclear. In the case of P. aeruginosa, the effector protein ExoS is central to limiting effector injection. ExoS is bifunctional, with an amino-terminal RhoGAP and a carboxy-terminal ADP-ribosyltransferase domain. We demonstrate that both domains are required to fully feedback inhibit effector injection. The RhoGAP-, but not the ADP-ribosyltransferase domain of the related effector protein ExoT also participates. Feedback inhibition does not involve translocator insertion nor pore-formation. Instead, feedback inhibition is due, in part, to a loss of the activating trigger for effector injection, and likely also decreased translocon stability. Surprisingly, feedback inhibition is abrogated in phagocytic cells. The lack of feedback inhibition in these cells requires phagocytic uptake of the bacteria, but cannot be explained through acidification of the phagosome or calcium limitation. Given that phagocytes are crucial for controlling P. aeruginosa infections, our data suggest that feedback inhibition allows P. aeruginosa to direct its effector arsenal against the cell types most damaging to its survival.
许多革兰氏阴性病原体使用 III 型分泌系统(T3SS)将效应蛋白注入宿主细胞,从而促进疾病的发生。许多 T3SS 的共同点是,效应蛋白的注入受到反馈抑制。反馈抑制的机制及其在发病机制中的作用尚不清楚。在铜绿假单胞菌的情况下,效应蛋白 ExoS 是限制效应蛋白注入的核心。ExoS 具有双重功能,氨基端 RhoGAP 和羧基端 ADP-核糖基转移酶结构域。我们证明这两个结构域都需要完全反馈抑制效应蛋白的注入。相关效应蛋白 ExoT 的 RhoGAP 结构域,但不是 ADP-核糖基转移酶结构域也参与其中。反馈抑制不涉及转运器插入或孔形成。相反,反馈抑制部分是由于效应蛋白注入的激活触发物丢失,并且可能还降低了转位器的稳定性。令人惊讶的是,吞噬细胞中的反馈抑制被废除。这些细胞中缺乏反馈抑制需要细菌被吞噬细胞摄取,但不能通过吞噬体酸化或钙限制来解释。鉴于吞噬细胞对于控制铜绿假单胞菌感染至关重要,我们的数据表明,反馈抑制允许铜绿假单胞菌将其效应器武器针对对其生存最具破坏性的细胞类型。