Host FIH-Mediated Asparaginyl Hydroxylation of TranslocatedLegionella pneumophilaEffectors.
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 11Legionella pneumophilatranslocated effectors, YopM ofYersinia, IpaH4.5 ofShigellaand an ankyrin protein ofRickettsia. Mass spectrometry analyses of the AnkB and AnkH effectors ofL. pneumophilaconfirm their asparaginyl hydroxylation. Consistent with localization of the AnkB effector to theLegionella-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 ofL. pneumophila. 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.
Department of Microbiology and Immunology, College of Medicine, University of Louisville Louisville, KY, USA.
Christopher
C
Price
Department of Medicine-Renal, College of Medicine, University of Louisville Louisville, KY, USA.
Michael
M
Merchant
Department of Microbiology and Immunology, College of Medicine, University of Louisville Louisville, KY, USA.
Snake
S
Jones
Department of Microbiology and Immunology, College of Medicine, University of Louisville Louisville, KY, USA.
Ashley
A
Best
Department of Microbiology and Immunology, College of Medicine, University of Louisville Louisville, KY, USA.
Juanita
J
Von Dwingelo
Department of Microbiology and Immunology, College of Medicine, University of LouisvilleLouisville, KY, USA; Center for Predictive Medicine, College of Medicine, University of LouisvilleLouisville, KY, USA.
Matthew B
MB
Lawrenz
Department of Microbiology and Molecular Genetics, Institute for Medical Research Israel-Canada (IMRIC), Faculty of Medicine, Hadassah Medical School, The Hebrew University of Jerusalem Jerusalem, Israel.
Nawsad
N
Alam
Department of Microbiology and Molecular Genetics, Institute for Medical Research Israel-Canada (IMRIC), Faculty of Medicine, Hadassah Medical School, The Hebrew University of Jerusalem Jerusalem, Israel.
Ora
O
Schueler-Furman
Department of Microbiology and Immunology, College of Medicine, University of LouisvilleLouisville, KY, USA; Center for Predictive Medicine, College of Medicine, University of LouisvilleLouisville, KY, USA.
Yousef A
YA
Kwaik
D020816:Amino Acid Motifs; D001216:Asparagine / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D016384:Consensus Sequence; D015964:Gene Expression Regulation, Bacterial; D054884:Host-Pathogen Interactions; D006801:Humans; D006900:Hydroxylation; D016952:Legionella pneumophila / Q000502:physiology*; D007877:Legionnaires' Disease / Q000378:metabolism* / Q000382:microbiology*; D050296:Microbial Viability; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000378:metabolism*; D009154:Mutation; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D012097:Repressor Proteins / Q000737:chemistry / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism
D001426:Bacterial Proteins; D012097:Repressor Proteins; D025801:Ubiquitin; D001216:Asparagine; D006899:Mixed Function Oxygenases; C439790:HIF1AN protein, human
AnkB; Dot/Icm; FIH; Legionella; ankyrin; asparagine hydroxylation; bacterial pathogenesis; hypoxia-inducible factor (HIF)