Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
Molecular Medicine Program, The Hospital for Sick Children, Toronto, Ontario, Canada.
J Biol Chem. 2023 Oct;299(10):105199. doi: 10.1016/j.jbc.2023.105199. Epub 2023 Sep 3.
Regulatory ATPase variant A (RavA) is a MoxR AAA+ protein that functions together with a partner protein termed von Willebrand factor type A interacting with AAA+ ATPase (ViaA). RavA-ViaA are functionally associated with anaerobic respiration in Escherichia coli through interactions with the fumarate reductase (Frd) electron transport complex. Through this association, RavA and ViaA modulate the activity of the Frd complex and, hence, are proposed to have chaperone-like activity. However, the functional role of RavA-ViaA in the cell is not yet well established. We had demonstrated that RavA-ViaA can sensitize E. coli cells to sublethal concentrations of the aminoglycoside class of antibiotics. Since Frd has been associated with bacterial persistence against antibiotics, the relationship of RavA-ViaA and Frd was explored within this context. Experiments performed here reveal a function of RavA-ViaA in bacterial persistence upon treatment with antibiotics through the association of the chaperone complex with Frd. As part of this work, the NMR structure of the N-terminal domain of ViaA was solved. The structure reveals a novel alpha helical fold, which we name the VAN fold, that has not been observed before. We show that this domain is required for the function of the chaperone complex. We propose that modulating the levels of RavA-ViaA could enhance the susceptibility of Gram-negative bacteria to antibiotics.
调节型 ATP 酶变体 A (RavA) 是一种 MoxR AAA+ 蛋白,与一种称为与 AAA+ATP 酶相互作用的血管性血友病因子 A 型 (ViaA) 的伴侣蛋白一起发挥作用。RavA-ViaA 通过与延胡索酸还原酶 (Frd) 电子传递复合物相互作用,在大肠杆菌的无氧呼吸中具有功能相关性。通过这种关联,RavA 和 ViaA 调节 Frd 复合物的活性,因此被认为具有伴侣样活性。然而,RavA-ViaA 在细胞中的功能作用尚未得到很好的确定。我们已经证明 RavA-ViaA 可以使大肠杆菌细胞对亚致死浓度的氨基糖苷类抗生素敏感。由于 Frd 与细菌对抗生素的持久性有关,因此在这种情况下探索了 RavA-ViaA 和 Frd 之间的关系。这里进行的实验揭示了 RavA-ViaA 通过伴侣复合物与 Frd 的关联,在抗生素处理时在细菌持久性中的功能。作为这项工作的一部分,解决了 ViaA 的 N 端结构域的 NMR 结构。该结构揭示了一种以前未观察到的新型α螺旋折叠,我们将其命名为 VAN 折叠。我们表明该结构域是伴侣复合物功能所必需的。我们提出,调节 RavA-ViaA 的水平可以增强革兰氏阴性菌对抗生素的敏感性。