Phenylpyrrole fungicides act on triosephosphate isomerase to induce methylglyoxal stress and alter hybrid histidine kinase activity
Phenylpyrrole fungicides act on triosephosphate isomerase to induce methylglyoxal stress and alter hybrid histidine kinase activity
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DOI：
10.1038/s41598-019-41564-9
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发表时间：
2019-03-25
影响因子：
4.6
通讯作者：
Klein, Bruce S.
中科院分区：
文献类型：
--
作者：
Brandhorst, T. Tristan;Kean, Iain R. L.;Klein, Bruce S.
咯菌腈是吡咯尼群的天然产物，是一种用于全世界农作物的强效杀菌剂。药物作用需要 III 族杂合组氨酸激酶 (HHK) 和 (HHK) 以及高渗透性甘油 (HOG) 途径的存在。我们报道该药物并不直接作用于 HHK，而是触发激酶转化为磷酸酶，从而使 Ypd1 去磷酸化，从而持续激活 HOG 信号传导。尽管如此，直接药物靶点仍然未知，作用方式也不清楚。在这里，我们在酿酒酵母中异源表达了 III 组 HHK，即二态性调节激酶 1 (Drk1)，以描述咯菌腈的靶点和作用。我们发现该药物会干扰磷酸三糖异构酶（TPI），导致甲基乙二醛（MG）的释放。 MG 激活 III 组 HHK，从而激活 HOG 途径。药物作用涉及 Drk1 半胱氨酸 392，因为 C392S 取代增加了体内耐药性。双甲酮治疗可逆转药物敏感性，表明 Drk1 在体内对醛应激有反应。咯菌腈处理引发胞质甲基乙二醛升高。同样，表达 Drk1 的酵母的甲基乙二醛处理与咯菌腈的处理相似。咯菌腈直接抑制 TPI，并使其在体外释放甲基乙二醛。因此，TPI 是苯基吡咯类杀菌剂的药物靶标，诱导 MG 升高，从而改变 HHK 活性，可能将激酶转化为作用于 Ypd1 的磷酸酶，从而触发 HOG 途径激活和真菌细胞死亡。
Fludioxonil, a natural product of pyrrolnitrin, is a potent fungicide used on crops worldwide. Drug action requires the presence of a group III hybrid histidine kinase (HHK) and the (HHK) and the high osmolarity glycerol (HOG) pathway. We have reported that the drug does not act directly on HHK, but triggers the conversion of the kinase to a phosphatase, which dephosphorylatesYpd1 to constitutively activate HOG signaling. Still, the direct drug target remains unknown and mode of action ill defined. Here, we heterologously expressed a group III HHK, dimorphism-regulating kinase 1 (Drk1) in Saccharomyces cerevisae to delineate fludioxonil's target and action. We show that the drug interferes with triosephosphate isomerase (TPI) causing release of methylglyoxal (MG). MG activates the group III HHK and thus the HOG pathway. Drug action involved Drk1 cysteine 392, as a C392S substitution increased drug resistance in vivo. Drug sensitivity was reversed by dimedone treatment, indicating Drk1 responds in vivo to an aldehydic stress. Fludioxonil treatment triggered elevated cytosolic methylglyoxal. Likewise, methylglyoxal treatment of Drk1-expressing yeast phenocopied treatment with fludioxonil. Fludioxonil directly inhibited TPI and also caused it to release methylglyoxal in vitro. Thus, TPI is a drug target of the phenylpyrrole class of fungicides, inducing elevated MG which alters HHK activity, likely converting the kinase to a phosphatase that acts on Ypd1 to trigger HOG pathway activation and fungal cell death.