The Legionella Pneumophila Metaeffector Lpg2505 (MesI) Regulates SidI-Mediated Translation Inhibition and Novel Glycosyl Hydrolase Activity

Overview

Journal Infect Immun

Publisher American Society for Microbiology

Specialty Allergy & Immunology

Date 2020 Mar 4

PMID 32122942

Citations 18

Authors

Ashley M Joseph

Adrienne E Pohl

Theodore J Ball

Troy G Abram

David K Johnson

Brian V Geisbrecht

Stephanie R Shames

Affiliations

Soon will be listed here.

Abstract

, the etiological agent of Legionnaires' disease, employs an arsenal of hundreds of Dot/Icm-translocated effector proteins to facilitate replication within eukaryotic phagocytes. Several effectors, called metaeffectors, function to regulate the activity of other Dot/Icm-translocated effectors during infection. The metaeffector Lpg2505 is essential for intracellular replication only when its cognate effector, SidI, is present. SidI is a cytotoxic effector that interacts with the host translation factor eEF1A and potently inhibits eukaryotic protein translation by an unknown mechanism. Here, we evaluated the impact of Lpg2505 on SidI-mediated phenotypes and investigated the mechanism of SidI function. We determined that Lpg2505 binds with nanomolar affinity to SidI and suppresses SidI-mediated inhibition of protein translation. SidI binding to eEF1A and Lpg2505 is not mutually exclusive, and the proteins bind distinct regions of SidI. We also discovered that SidI possesses GDP-dependent glycosyl hydrolase activity and that this activity is regulated by Lpg2505. We have therefore renamed Lpg2505 MesI (taeffector of id). This work reveals novel enzymatic activity for SidI and provides insight into how intracellular replication of is regulated by a metaeffector.

Citing Articles

The effector PieF modulates mRNA stability through association with eukaryotic CCR4-NOT.

Mount H, Urbanus M, Zangari F, Gingras A, Ensminger A mSphere. 2024; 10(1):e0089124.

PMID: 39699231 PMC: 11774319. DOI: 10.1128/msphere.00891-24.

, a Rosetta stone to understanding bacterial pathogenesis.

Romanov K, OConnor T J Bacteriol. 2024; 206(12):e0032424.

PMID: 39636264 PMC: 11656745. DOI: 10.1128/jb.00324-24.

Global atlas of predicted functional domains in Legionella pneumophila Dot/Icm translocated effectors.

Patel D, Stogios P, Jaroszewski L, Urbanus M, Sedova M, Semper C Mol Syst Biol. 2024; 21(1):59-89.

PMID: 39562741 PMC: 11696984. DOI: 10.1038/s44320-024-00076-z.

A comprehensive two-hybrid analysis to explore the effector-effector interactome.

Mount H, Urbanus M, Sheykhkarimli D, Cote A, Laval F, Coppin G mSystems. 2024; 9(12):e0100424.

PMID: 39526800 PMC: 11651115. DOI: 10.1128/msystems.01004-24.

InSeq analysis of defined libraries identifies a transporter-encoding gene cluster important for intracellular replication in mammalian hosts.

Moss C, Roy C mBio. 2024; 15(11):e0195524.

PMID: 39365064 PMC: 11559062. DOI: 10.1128/mbio.01955-24.

References

Kelley L, Mezulis S, Yates C, Wass M, Sternberg M . The Phyre2 web portal for protein modeling, prediction and analysis. Nat Protoc. 2015; 10(6):845-58. PMC: 5298202. DOI: 10.1038/nprot.2015.053. View

Tzivelekidis T, Jank T, Pohl C, Schlosser A, Rospert S, Knudsen C . Aminoacyl-tRNA-charged eukaryotic elongation factor 1A is the bona fide substrate for Legionella pneumophila effector glucosyltransferases. PLoS One. 2012; 6(12):e29525. PMC: 3245282. DOI: 10.1371/journal.pone.0029525. View

Geisbrecht B, Bouyain S, Pop M . An optimized system for expression and purification of secreted bacterial proteins. Protein Expr Purif. 2005; 46(1):23-32. DOI: 10.1016/j.pep.2005.09.003. View

Jeong K, Sexton J, Vogel J . Spatiotemporal regulation of a Legionella pneumophila T4SS substrate by the metaeffector SidJ. PLoS Pathog. 2015; 11(3):e1004695. PMC: 4361747. DOI: 10.1371/journal.ppat.1004695. View

Yang J, Zhang Y . I-TASSER server: new development for protein structure and function predictions. Nucleic Acids Res. 2015; 43(W1):W174-81. PMC: 4489253. DOI: 10.1093/nar/gkv342. View

Wang Y, Shi M, Feng H, Zhu Y, Liu S, Gao A . Structural Insights into Non-canonical Ubiquitination Catalyzed by SidE. Cell. 2018; 173(5):1231-1243.e16. DOI: 10.1016/j.cell.2018.04.023. View

Price C, Al-Quadan T, Santic M, Rosenshine I, Abu Kwaik Y . Host proteasomal degradation generates amino acids essential for intracellular bacterial growth. Science. 2011; 334(6062):1553-7. DOI: 10.1126/science.1212868. View

Belyi I, Popoff M, Cianciotto N . Purification and characterization of a UDP-glucosyltransferase produced by Legionella pneumophila. Infect Immun. 2002; 71(1):181-6. PMC: 143419. DOI: 10.1128/IAI.71.1.181-186.2003. View

Bhogaraju S, Bonn F, Mukherjee R, Adams M, Pfleiderer M, Galej W . Inhibition of bacterial ubiquitin ligases by SidJ-calmodulin catalysed glutamylation. Nature. 2019; 572(7769):382-386. PMC: 6715450. DOI: 10.1038/s41586-019-1440-8. View

10.

Belyi Y, Tabakova I, Stahl M, Aktories K . Lgt: a family of cytotoxic glucosyltransferases produced by Legionella pneumophila. J Bacteriol. 2008; 190(8):3026-35. PMC: 2293231. DOI: 10.1128/JB.01798-07. View

11.

Yan L, Cerny R, Cirillo J . Evidence that hsp90 is involved in the altered interactions of Acanthamoeba castellanii variants with bacteria. Eukaryot Cell. 2004; 3(3):567-78. PMC: 420136. DOI: 10.1128/EC.3.3.567-578.2004. View

12.

Sethi M, Buettner F, Ashikov A, Bakker H . In vitro assays of orphan glycosyltransferases and their application to identify Notch xylosyltransferases. Methods Mol Biol. 2013; 1022:307-20. DOI: 10.1007/978-1-62703-465-4_23. View

13.

Berger K, Isberg R . Two distinct defects in intracellular growth complemented by a single genetic locus in Legionella pneumophila. Mol Microbiol. 1993; 7(1):7-19. DOI: 10.1111/j.1365-2958.1993.tb01092.x. View

14.

Gan N, Zhen X, Liu Y, Xu X, He C, Qiu J . Regulation of phosphoribosyl ubiquitination by a calmodulin-dependent glutamylase. Nature. 2019; 572(7769):387-391. PMC: 6855250. DOI: 10.1038/s41586-019-1439-1. View

15.

Urbanus M, Quaile A, Stogios P, Morar M, Rao C, Di Leo R . Diverse mechanisms of metaeffector activity in an intracellular bacterial pathogen, Legionella pneumophila. Mol Syst Biol. 2016; 12(12):893. PMC: 5199130. DOI: 10.15252/msb.20167381. View

16.

Heidtman M, Chen E, Moy M, Isberg R . Large-scale identification of Legionella pneumophila Dot/Icm substrates that modulate host cell vesicle trafficking pathways. Cell Microbiol. 2008; 11(2):230-48. PMC: 2744955. DOI: 10.1111/j.1462-5822.2008.01249.x. View

17.

Belyi Y, Niggeweg R, Opitz B, Vogelsgesang M, Hippenstiel S, Wilm M . Legionella pneumophila glucosyltransferase inhibits host elongation factor 1A. Proc Natl Acad Sci U S A. 2006; 103(45):16953-8. PMC: 1636560. DOI: 10.1073/pnas.0601562103. View

18.

Lu W, Du J, Stahl M, Tzivelekidis T, Belyi Y, Gerhardt S . Structural basis of the action of glucosyltransferase Lgt1 from Legionella pneumophila. J Mol Biol. 2009; 396(2):321-31. DOI: 10.1016/j.jmb.2009.11.044. View

19.

Shen X, Banga S, Liu Y, Xu L, Gao P, Shamovsky I . Targeting eEF1A by a Legionella pneumophila effector leads to inhibition of protein synthesis and induction of host stress response. Cell Microbiol. 2009; 11(6):911-26. PMC: 2967282. DOI: 10.1111/j.1462-5822.2009.01301.x. View

20.

Guo Z, Stephenson R, Qiu J, Zheng S, Luo Z . A Legionella effector modulates host cytoskeletal structure by inhibiting actin polymerization. Microbes Infect. 2013; 16(3):225-36. PMC: 3965633. DOI: 10.1016/j.micinf.2013.11.007. View