Unité de Microbiologie Structurale, Institut Pasteur, CNRS UMR 3528 & Université Paris Diderot, 75724 Paris Cedex 15, France.
Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR), Ocampo y Esmeralda, S2002LRK, Rosario, Argentina.
Nucleic Acids Res. 2019 Apr 23;47(7):3795-3810. doi: 10.1093/nar/gkz112.
Upon triggering by their inducer, signal transduction ATPases with numerous domains (STANDs), initially in monomeric resting forms, multimerize into large hubs that activate target macromolecules. This process requires conversion of the STAND conserved core (the NOD) from a closed form encasing an ADP molecule to an ATP-bound open form prone to multimerize. In the absence of inducer, autoinhibitory interactions maintain the NOD closed. In particular, in resting STAND proteins with an LRR- or WD40-type sensor domain, the latter establishes interactions with the NOD that are disrupted in the multimerization-competent forms. Here, we solved the first crystal structure of a STAND with a tetratricopeptide repeat sensor domain, PH0952 from Pyrococcus horikoshii, revealing analogous NOD-sensor contacts. We use this structural information to experimentally demonstrate that similar interactions also exist in a PH0952 homolog, the MalT STAND archetype, and actually contribute to the MalT autoinhibition in vitro and in vivo. We propose that STAND activation occurs by stepwise release of autoinhibitory contacts coupled to the unmasking of inducer-binding determinants. The MalT example suggests that STAND weak autoinhibitory interactions could assist the binding of inhibitory proteins by placing in register inhibitor recognition elements born by two domains.
在被其诱导剂触发后,具有众多结构域的信号转导 ATP 酶(STAND)最初以单体静息形式存在,会聚合形成大的中心体,从而激活靶标大分子。这一过程需要将 STAND 保守核心(NOD)从包含 ADP 分子的封闭形式转换为易于聚合的 ATP 结合的开放形式。在没有诱导剂的情况下,自动抑制相互作用会保持 NOD 关闭。特别是在具有 LRR 或 WD40 型传感器结构域的静息 STAND 蛋白中,后者与 NOD 建立相互作用,而在具有多聚化能力的形式中,这种相互作用会被破坏。在这里,我们解析了第一个具有四肽重复传感器结构域的 STAND 的晶体结构,即来自 Pyrococcus horikoshii 的 PH0952,揭示了类似的 NOD-传感器接触。我们利用这些结构信息,通过实验证明,类似的相互作用也存在于 PH0952 同源物,MalT STAND 原型中,并且实际上有助于 MalT 在体外和体内的自动抑制。我们提出,STAND 的激活是通过逐步释放自动抑制相互作用与诱导剂结合决定因素的暴露来实现的。MalT 的例子表明,STAND 较弱的自动抑制相互作用可以通过将两个结构域上的抑制剂识别元件置于适当的位置,来帮助抑制蛋白的结合。