Price Christopher, Merchant Michael, Jones Snake, Best Ashley, Von Dwingelo Juanita, Lawrenz Matthew B, Alam Nawsad, Schueler-Furman Ora, Kwaik Yousef A
Department of Microbiology and Immunology, College of Medicine, University of Louisville Louisville, KY, USA.
Department of Medicine-Renal, College of Medicine, University of Louisville Louisville, KY, USA.
Front Cell Infect Microbiol. 2017 Mar 6;7:54. doi: 10.3389/fcimb.2017.00054. eCollection 2017.
FIH-mediated post-translational modification through asparaginyl hydroxylation of eukaryotic proteins impacts regulation of protein-protein interaction. We have identified the FIH recognition motif in 11 translocated effectors, YopM of , IpaH4.5 of and an ankyrin protein of . Mass spectrometry analyses of the AnkB and AnkH effectors of confirm their asparaginyl hydroxylation. Consistent with localization of the AnkB effector to the -containing vacuole (LCV) membrane and its modification by FIH, our data show that FIH and its two interacting proteins, Mint3 and MT1-MMP are acquired by the LCV in a Dot/Icm type IV secretion-dependent manner. Chemical inhibition or RNAi-mediated knockdown of FIH promotes LCV-lysosomes fusion, diminishes decoration of the LCV with polyubiquitinated proteins, and abolishes intra-vacuolar replication of . These data show acquisition of the host FIH by a pathogen-containing vacuole and that asparaginyl-hydroxylation of translocated effectors is indispensable for their function.
因子抑制因子（FIH）介导的真核蛋白质天冬酰胺羟基化的翻译后修饰影响蛋白质-蛋白质相互作用的调控。我们在11种转运效应蛋白中鉴定出了FIH识别基序，包括耶尔森氏菌的YopM、痢疾杆菌的IpaH4.5和军团菌的一种锚蛋白。对嗜肺军团菌的AnkB和AnkH效应蛋白的质谱分析证实了它们的天冬酰胺羟基化。与AnkB效应蛋白定位于含军团菌液泡（LCV）膜并被FIH修饰一致，我们的数据表明，FIH及其两个相互作用蛋白Mint3和MT1-MMP以Dot/Icm IV型分泌依赖的方式被LCV获取。化学抑制或RNA干扰介导的FIH敲低促进LCV-溶酶体融合，减少LCV上多聚泛素化蛋白的修饰，并消除嗜肺军团菌在液泡内的复制。这些数据表明含病原体液泡获取了宿主FIH，并且转运效应蛋白的天冬酰胺羟基化对其功能不可或缺。
