CYRI/FAM49B negatively regulates RAC1-driven cytoskeletal remodelling and protects against bacterial infection.
Actins
/ metabolism
Animals
Bacterial Infections
/ metabolism
Bacterial Load
Bacterial Proteins
/ genetics
Cytoskeleton
/ genetics
Disease Resistance
/ genetics
HEK293 Cells
HeLa Cells
Host-Pathogen Interactions
Humans
Intracellular Signaling Peptides and Proteins
/ genetics
Listeria monocytogenes
/ metabolism
Macrophages
/ microbiology
Mice
Mitochondrial Proteins
/ genetics
Mutation
Mycobacterium tuberculosis
/ metabolism
Phagocytosis
Protein Binding
Salmonella typhimurium
/ metabolism
Signal Transduction
Survival Analysis
rac1 GTP-Binding Protein
/ metabolism
Journal
Nature microbiology
ISSN: 2058-5276
Titre abrégé: Nat Microbiol
Pays: England
ID NLM: 101674869
Informations de publication
Date de publication:
09 2019
09 2019
Historique:
received:
26
04
2018
accepted:
08
05
2019
pubmed:
10
7
2019
medline:
22
1
2020
entrez:
10
7
2019
Statut:
ppublish
Résumé
Salmonella presents a global public health concern. Central to Salmonella pathogenicity is an ability to subvert host defences through strategically targeting host proteins implicated in restricting infection. Therefore, to gain insight into the host-pathogen interactions governing Salmonella infection, we performed an in vivo genome-wide mutagenesis screen to uncover key host defence proteins. This revealed an uncharacterized role of CYRI (FAM49B) in conferring host resistance to Salmonella infection. We show that CYRI binds to the small GTPase RAC1 through a conserved domain present in CYFIP proteins, which are known RAC1 effectors that stimulate actin polymerization. However, unlike CYFIP proteins, CYRI negatively regulates RAC1 signalling, thereby attenuating processes such as macropinocytosis, phagocytosis and cell migration. This enables CYRI to counteract Salmonella at various stages of infection, including bacterial entry into non-phagocytic and phagocytic cells as well as phagocyte-mediated bacterial dissemination. Intriguingly, to dampen its effects, the bacterial effector SopE, a RAC1 activator, selectively targets CYRI following infection. Together, this outlines an intricate host-pathogen signalling interplay that is crucial for determining bacterial fate. Notably, our study also outlines a role for CYRI in restricting infection mediated by Mycobacterium tuberculosis and Listeria monocytogenes. This provides evidence implicating CYRI cellular functions in host defence beyond Salmonella infection.
Identifiants
pubmed: 31285585
doi: 10.1038/s41564-019-0484-8
pii: 10.1038/s41564-019-0484-8
doi:
Substances chimiques
Actins
0
Bacterial Proteins
0
CYRIB protein, human
0
Intracellular Signaling Peptides and Proteins
0
Mitochondrial Proteins
0
SopE protein, Salmonella
0
rac1 GTP-Binding Protein
EC 3.6.5.2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1516-1531Références
Kirk, M. D. et al. World Health Organization estimates of the global and regional disease burden of 22 foodborne bacterial, protozoal, and viral diseases, 2010: a data synthesis. PLoS Med. 12, e1001921 (2015).
doi: 10.1371/journal.pmed.1001921
Majowicz, S. E. et al. The global burden of nontyphoidal Salmonella gastroenteritis. Clin. Infect. Dis. 50, 882–889 (2010).
doi: 10.1086/650733
Prestinaci, F., Pezzotti, P. & Pantosti, A. Antimicrobial resistance: a global multifaceted phenomenon. Pathog. Glob. Health 109, 309–318 (2015).
doi: 10.1179/2047773215Y.0000000030
LaRock, D. L., Chaudhary, A. & Miller, S. I. Salmonellae interactions with host processes. Nat. Rev. Microbiol. 13, 191–205 (2015).
doi: 10.1038/nrmicro3420
Schlumberger, M. C. & Hardt, W. D. Salmonella type III secretion effectors: pulling the host cell’s strings. Curr. Opin. Microbiol. 9, 46–54 (2006).
doi: 10.1016/j.mib.2005.12.006
Byndloss, M. X., Rivera-Chavez, F., Tsolis, R. M. & Baumler, A. J. How bacterial pathogens use type III and type IV secretion systems to facilitate their transmission. Curr. Opin. Microbiol. 35, 1–7 (2017).
doi: 10.1016/j.mib.2016.08.007
Guiney, D. G. & Lesnick, M. Targeting of the actin cytoskeleton during infection by Salmonella strains. Clin. Immunol. 114, 248–255 (2005).
doi: 10.1016/j.clim.2004.07.014
Hardt, W. D., Chen, L. M., Schuebel, K. E., Bustelo, X. R. & Galan, J. E. S. typhimurium encodes an activator of Rho GTPases that induces membrane ruffling and nuclear responses in host cells. Cell 93, 815–826 (1998).
doi: 10.1016/S0092-8674(00)81442-7
Orchard, R. C. & Alto, N. M. Mimicking GEFs: a common theme for bacterial pathogens. Cell. Microbiol. 14, 10–18 (2012).
doi: 10.1111/j.1462-5822.2011.01703.x
Humphreys, D., Davidson, A., Hume, P. J. & Koronakis, V. Salmonella virulence effector SopE and host GEF ARNO cooperate to recruit and activate WAVE to trigger bacterial invasion. Cell Host Microbe 11, 129–139 (2012).
doi: 10.1016/j.chom.2012.01.006
Gautreau, A. et al. Purification and architecture of the ubiquitous Wave complex. Proc. Natl Acad. Sci. USA 101, 4379–4383 (2004).
doi: 10.1073/pnas.0400628101
Eden, S., Rohatgi, R., Podtelejnikov, A. V., Mann, M. & Kirschner, M. W. Mechanism of regulation of WAVE1-induced actin nucleation by Rac1 and Nck. Nature 418, 790–793 (2002).
doi: 10.1038/nature00859
Ismail, A. M., Padrick, S. B., Chen, B., Umetani, J. & Rosen, M. K. The WAVE regulatory complex is inhibited. Nat. Struct. Mol. Biol. 16, 561–563 (2009).
doi: 10.1038/nsmb.1587
Chen, Z. et al. Structure and control of the actin regulatory WAVE complex. Nature 468, 533–538 (2010).
doi: 10.1038/nature09623
Chen, B. et al. Rac1 GTPase activates the WAVE regulatory complex through two distinct binding sites. eLife 6, e29795 (2017).
doi: 10.7554/eLife.29795
Francis, C. L., Ryan, T. A., Jones, B. D., Smith, S. J. & Falkow, S. Ruffles induced by Salmonella and other stimuli direct macropinocytosis of bacteria. Nature 364, 639–642 (1993).
doi: 10.1038/364639a0
Steele-Mortimer, O. The Salmonella-containing vacuole: moving with the times. Curr. Opin. Microbiol. 11, 38–45 (2008).
doi: 10.1016/j.mib.2008.01.002
Ham, H., Sreelatha, A. & Orth, K. Manipulation of host membranes by bacterial effectors. Nat. Rev. Microbiol. 9, 635–646 (2011).
doi: 10.1038/nrmicro2602
Schleker, S. et al. The current Salmonella-host interactome. Proteom. Clin. Appl. 6, 117–133 (2012).
doi: 10.1002/prca.201100083
Mostowy, S. & Shenoy, A. R. The cytoskeleton in cell-autonomous immunity: structural determinants of host defence. Nat. Rev. Immunol. 15, 559–573 (2015).
doi: 10.1038/nri3877
Behnsen, J., Perez-Lopez, A., Nuccio, S. P. & Raffatellu, M. Exploiting host immunity: the Salmonella paradigm. Trends Immunol. 36, 112–120 (2015).
doi: 10.1016/j.it.2014.12.003
Worley, M. J., Nieman, G. S., Geddes, K. & Heffron, F. Salmonella typhimurium disseminates within its host by manipulating the motility of infected cells. Proc. Natl Acad. Sci. USA 103, 17915–17920 (2006).
doi: 10.1073/pnas.0604054103
Ohl, M. E. & Miller, S. I. Salmonella: a model for bacterial pathogenesis. Annu. Rev. Med. 52, 259–274 (2001).
doi: 10.1146/annurev.med.52.1.259
Fabrega, A. & Vila, J. Salmonella enterica serovar Typhimurium skills to succeed in the host: virulence and regulation. Clin. Microbiol. Rev. 26, 308–341 (2013).
doi: 10.1128/CMR.00066-12
Vazquez-Torres, A. et al. Extraintestinal dissemination of Salmonella by CD18-expressing phagocytes. Nature 401, 804–808 (1999).
doi: 10.1038/44593
Hendriksen, R. S. et al. Global monitoring of Salmonella serovar distribution from the world health organization global foodborne infections network country data bank: results of quality assured laboratories from 2001 to 2007. Foodborne Pathog. Dis. 8, 887–900 (2011).
doi: 10.1089/fpd.2010.0787
Soding, J., Biegert, A. & Lupas, A. N. The HHpred interactive server for protein homology detection and structure prediction. Nucleic Acids Res. 33, W244–W248 (2005).
doi: 10.1093/nar/gki408
Michiels, F., Habets, G. G., Stam, J. C., van der Kammen, R. A. & Collard, J. G. A role for Rac in Tiam1-induced membrane ruffling and invasion. Nature 375, 338–340 (1995).
doi: 10.1038/375338a0
Krauthammer, M. et al. Exome sequencing identifies recurrent somatic RAC1 mutations in melanoma. Nat. Genet. 44, 1006–1014 (2012).
doi: 10.1038/ng.2359
Pollard, T. D. & Cooper, J. A. Actin, a central player in cell shape and movement. Science 326, 1208–1212 (2009).
doi: 10.1126/science.1175862
Spector, I., Shochet, N. R., Kashman, Y. & Groweiss, A. Latrunculins: novel marine toxins that disrupt microfilament organization in cultured cells. Science 219, 493–495 (1983).
doi: 10.1126/science.6681676
May, R. C. & Machesky, L. M. Phagocytosis and the actin cytoskeleton. J. Cell Sci. 114, 1061–1077 (2001).
pubmed: 11228151
Krause, M. & Gautreau, A. Steering cell migration: lamellipodium dynamics and the regulation of directional persistence. Nat. Rev. Mol. Cell Biol. 15, 577–590 (2014).
doi: 10.1038/nrm3861
Wijburg, O. L., Simmons, C. P., van Rooijen, N. & Strugnell, R. A. Dual role for macrophages in vivo in pathogenesis and control of murine Salmonella enterica var. Typhimurium infections. Eur. J. Immunol. 30, 944–953 (2000).
doi: 10.1002/1521-4141(200003)30:3<944::AID-IMMU944>3.0.CO;2-1
Richter-Dahlfors, A., Buchan, A. M. & Finlay, B. B. Murine salmonellosis studied by confocal microscopy: Salmonella typhimurium resides intracellularly inside macrophages and exerts a cytotoxic effect on phagocytes in vivo. J. Exp. Med. 186, 569–580 (1997).
doi: 10.1084/jem.186.4.569
Lin, D. C. & Lin, S. Actin polymerization induced by a motility-related high-affinity cytochalasin binding complex from human erythrocyte membrane. Proc. Natl Acad. Sci. USA 76, 2345–2349 (1979).
doi: 10.1073/pnas.76.5.2345
Campa, C. C. et al. Rac signal adaptation controls neutrophil mobilization from the bone marrow. Sci. Signal. 9, ra124 (2016).
doi: 10.1126/scisignal.aah5882
Pankov, R. et al. A Rac switch regulates random versus directionally persistent cell migration. J. Cell Biol. 170, 793–802 (2005).
doi: 10.1083/jcb.200503152
Schlumberger, M. C. et al. Amino acids of the bacterial toxin SopE involved in G nucleotide exchange on Cdc42. J. Biol. Chem. 278, 27149–27159 (2003).
doi: 10.1074/jbc.M302475200
Fiskin, E., Bionda, T., Dikic, I. & Behrends, C. Global analysis of host and bacterial ubiquitinome in response to Salmonella Typhimurium infection. Mol. Cell 62, 967–981 (2016).
doi: 10.1016/j.molcel.2016.04.015
de Chastellier, C. & Berche, P. Fate of Listeria monocytogenes in murine macrophages: evidence for simultaneous killing and survival of intracellular bacteria. Infect. Immun. 62, 543–553 (1994).
pubmed: 8300212
pmcid: 186140
Cossart, P. & Lebreton, A. A trip in the “New Microbiology” with the bacterial pathogen Listeria monocytogenes. FEBS Lett. 588, 2437–2445 (2014).
doi: 10.1016/j.febslet.2014.05.051
Rajaram, M. V. S. et al. M. tuberculosis-initiated human mannose receptor signaling regulates macrophage recognition and vesicle trafficking by FcRγ-Chain, Grb2, and SHP-1. Cell Rep. 21, 126–140 (2017).
doi: 10.1016/j.celrep.2017.09.034
Ireton, K., Rigano, L. A. & Dowd, G. C. Role of host GTPases in infection by Listeria monocytogenes. Cell Microbiol. 16, 1311–1320 (2014).
doi: 10.1111/cmi.12324
Shang, W. et al. Genome-wide CRISPR screen identifies FAM49B as a key regulator of actin dynamics and T cell activation. Proc. Natl Acad. Sci. USA 115, E4051–E4060 (2018).
doi: 10.1073/pnas.1801340115
Fort, L. et al. Fam49/CYRI interacts with Rac1 and locally suppresses protrusions. Nat. Cell Biol. 20, 1159–1171 (2018).
doi: 10.1038/s41556-018-0198-9
Queval, C. J., Brosch, R. & Simeone, R. The macrophage: a disputed fortress in the battle against Mycobacterium tuberculosis. Front. Microbiol. 8, 2284 (2017).
doi: 10.3389/fmicb.2017.02284
Haney, M. S. et al. Identification of phagocytosis regulators using magnetic genome-wide CRISPR screens. Nat. Genet. 50, 1716–1727 (2018).
doi: 10.1038/s41588-018-0254-1
Drevets, D. A. Dissemination of Listeria monocytogenes by infected phagocytes. Infect. Immun. 67, 3512–3517 (1999).
pubmed: 10377133
pmcid: 116538
Maculins, T., Fiskin, E., Bhogaraju, S. & Dikic, I. Bacteria-host relationship: ubiquitin ligases as weapons of invasion. Cell Res. 26, 499–510 (2016).
Roy, M. F. et al. Pyruvate kinase deficiency confers susceptibility to Salmonella typhimurium infection in mice. J. Exp. Med. 204, 2949–2961 (2007).
doi: 10.1084/jem.20062606
Barthel, M. et al. Pretreatment of mice with streptomycin provides a Salmonella enterica serovar Typhimurium colitis model that allows analysis of both pathogen and host. Infect. Immun. 71, 2839–2858 (2003).
doi: 10.1128/IAI.71.5.2839-2858.2003
Eva, M. M. et al. Altered IFN-γ-mediated immunity and transcriptional expression patterns in N-ethyl-N-nitrosourea-induced STAT4 mutants confer susceptibility to acute typhoid-like disease. J. Immunol. 192, 259–270 (2014).
doi: 10.4049/jimmunol.1301370
Marei, H. et al. Differential Rac1 signalling by guanine nucleotide exchange factors implicates FLII in regulating Rac1-driven cell migration. Nat. Commun. 7, 10664 (2016).
doi: 10.1038/ncomms10664
Benard, V., Bohl, B. P. & Bokoch, G. M. Characterization of Rac and Cdc42 activation in chemoattractant-stimulated human neutrophils using a novel assay for active GTPases. J. Biol. Chem. 274, 13198–13204 (1999).
doi: 10.1074/jbc.274.19.13198
Marei, H., Carpy, A., Macek, B. & Malliri, A. Proteomic analysis of Rac1 signaling regulation by guanine nucleotide exchange factors. Cell Cycle 15, 1961–1974 (2016).
doi: 10.1080/15384101.2016.1183852
Karlinsey, J. E. λ-Red genetic engineering in Salmonella enterica serovar Typhimurium. Methods Enzym. 421, 199–209 (2007).
doi: 10.1016/S0076-6879(06)21016-4
Skarnes, W. C. et al. A conditional knockout resource for the genome-wide study of mouse gene function. Nature 474, 337–342 (2011).
doi: 10.1038/nature10163
Fortin, A. et al. Recombinant congenic strains derived from A/J and C57BL/6J: a tool for genetic dissection of complex traits. Genomics 74, 21–35 (2001).
doi: 10.1006/geno.2001.6528
Cunnington, A. J., de Souza, J. B., Walther, M. & Riley, E. M. Malaria impairs resistance to Salmonella through heme- and heme oxygenase-dependent dysfunctional granulocyte mobilization. Nat. Med. 18, 120–127 (2011).
doi: 10.1038/nm.2601
Sixt, M. & Lammermann, T. In vitro analysis of chemotactic leukocyte migration in 3D environments. Methods Mol. Biol. 769, 149–165 (2011).
doi: 10.1007/978-1-61779-207-6_11