AUTHOR=Price Christopher , Merchant Michael , Jones Snake , Best Ashley , Von Dwingelo Juanita , Lawrenz Matthew B. , Alam Nawsad , Schueler-Furman Ora , Kwaik Yousef A. 

TITLE=Host FIH-Mediated Asparaginyl Hydroxylation of Translocated Legionella pneumophila Effectors

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=7

YEAR=2017

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2017.00054

DOI=10.3389/fcimb.2017.00054

ISSN=2235-2988

ABSTRACT=<p>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 <italic>Legionella pneumophila</italic> translocated effectors, YopM of <italic>Yersinia</italic>, IpaH4.5 of <italic>Shigella</italic> and an ankyrin protein of <italic>Rickettsia</italic>. Mass spectrometry analyses of the AnkB and AnkH effectors of <italic>L. pneumophila</italic> confirm their asparaginyl hydroxylation. Consistent with localization of the AnkB effector to the <italic>Legionella</italic>-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 <italic>L. pneumophila</italic>. 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.</p>