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Inhibition of the signal transduction through the AtoSC system by histidine kinase inhibitors in Escherichia coli

机译:组氨酸激酶抑制剂在大肠杆菌中抑制通过AtoSC系统的信号转导

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AtoSC two-component system participates in many indispensable processes of Escherichia coli. We report here that the AtoSC signal transduction is inhibited by established histidine kinase inhibitors. Closantel, RWJ-49815 and TNP-ATP belonging to different chemical classes of inhibitors, abrogated the in vitro AtoS kinase autophosphorylation. However, when AtoS was embedded in the membrane fractions, higher inhibitor concentrations were required for total inhibition. When AtoS interacted with AtoC forming complex, the intrinsic histidine kinase was protected by the response regulator, requiring increased inhibitors concentrations for partially AtoS autophosphorylation reduction. The inhibitors exerted an additional function on AtoSC, blocking the phosphotransfer from AtoS to AtoC, without however, affecting AtoC~P dephosphorylation. Their in vivo consequences through the AtoSC inhibition were elucidated on atoDAEB operon expression, which was inhibited only in AtoSC-expressing bacteria where AtoSC was induced by acetoacetate or spermidine. The inhibitor effects were extended on the AtoSC regulatory role on cPHB [complexed poly-(R)-3-hydroxybutyrate] biosynthesis. cPHB was decreased upon the blockers only in acetoacetate-induced AtoSC-expressing cells. Increased ATP amounts during bacterial growth reversed the inhibitory TNP-ATP-mediated effect on cPHB. The alteration of pivotal E. coli processes as an outcome of AtoSC inhibition, establish this system as a target of two-component systems inhibitors.
机译:AtoSC两组分系统参与大肠杆菌的许多必不可少的过程。我们在这里报告,AtoSC信号转导被建立的组氨酸激酶抑制剂抑制。属于不同化学类别的抑制剂的Closantel,RWJ-49815和TNP-ATP废除了体外AtoS激酶的自磷酸化作用。但是,当AtoS嵌入膜部分时,需要更高的抑制剂浓度才能完全抑制。当AtoS与AtoC形成复合物相互作用时,内在的组氨酸激酶受到反应调节剂的保护,需要增加抑制剂浓度以部分降低AtoS自磷酸化作用。抑制剂在AtoSC上发挥了额外的功能,阻止了从AtoS到AtoC的磷酸转移,但不影响AtoC〜P的去磷酸化。通过atoDAEB操纵子表达阐明了它们通过AtoSC抑制的体内结果,该表达仅在表达AtoSC的细菌中被抑制,其中AtoSC由乙酰乙酸或亚精胺诱导。抑制剂的作用扩展了对cPHB [复杂的聚-(R)-3-羟基丁酸酯]生物合成的AtoSC调节作用。 cPHB仅在乙酰乙酸诱导的表达AtoSC的细胞中在阻断剂后降低。细菌生长过程中增加的ATP量逆转了TNP-ATP介导的对cPHB的抑制作用。作为AtoSC抑制作用的结果,关键性大肠杆菌过程的改变使该系统成为两组分系统抑制剂的靶标。

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