Live Imaging of Mycobacterium marinum Infection in Dictyostelium discoideum

2015 ◽  
pp. 369-385 ◽  
Author(s):  
Caroline Barisch ◽  
Ana T. López-Jiménez ◽  
Thierry Soldati
Keyword(s):  
Dictyostelium Discoideum ◽  
Live Imaging ◽  
Mycobacterium Marinum

scholarly journals Disruption of vacuolin microdomains in the host Dictyostelium discoideum increases resistance to Mycobacterium marinum-induced membrane damage and infection

2021 ◽  
Author(s):  
Cristina Bosmani ◽  
Angélique Perret ◽  
Florence Leuba ◽  
Aurélie Guého ◽  
Nabil Hanna ◽  
...  
Keyword(s):  
Dictyostelium Discoideum ◽  
Membrane Damage ◽  
Mycobacterium Marinum ◽  
Vesicular Trafficking ◽  
Membrane Microdomains ◽  
Membrane Association ◽  
Phagosome Maturation ◽  
Closely Related Species ◽  
Important Host ◽  
And Function

Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, manipulates the host phagosome maturation pathway to replicate intracellularly. Mycobacterium marinum, a closely-related species, and Dictyostelium discoideum, a social amoeba and alternative phagocytic host, have been used as models to study host-pathogen interactions occurring during mycobacterial infections. Vacuolins, functional homologues of the mammalian flotillins, organize membrane microdomains and play a role in vesicular trafficking. Various pathogens have been reported to manipulate their membrane association and function. During infection of D. discoideum with M. marinum, Vacuolin C was specifically and highly induced and all three vacuolin isoforms were enriched at the mycobacteria-containing-vacuole (MCV). In addition, absence of vacuolins reduced escape from the MCV and conferred resistance to M. marinum infection. Moreover, ESAT-6, the membrane-disrupting virulence factor of M. marinum, was less associated with membranes when vacuolins were absent. Together, these results suggest that vacuolins are important host factors that are manipulated by mycobacteria to inflict membrane damage and escape from their compartment.

Novel Single-Cell and High-Throughput Microscopy Techniques to Monitor Dictyostelium discoideum–Mycobacterium marinum Infection Dynamics

2021 ◽  
pp. 183-203
Author(s):  
Manon Mottet ◽  
Cristina Bosmani ◽  
Nabil Hanna ◽  
Jahn Nitschke ◽  
Louise H. Lefrançois ◽  
...  
Keyword(s):  
Single Cell ◽  
Dictyostelium Discoideum ◽  
High Throughput ◽  
Mycobacterium Marinum ◽  
Infection Dynamics ◽  
Microscopy Techniques

scholarly journals Intracellular Replication of Mycobacterium marinum within Dictyostelium discoideum: Efficient Replication in the Absence of Host Coronin

2003 ◽  
Vol 71 (6) ◽  
pp. 3578-3586 ◽  
Author(s):  
Jonathan M. Solomon ◽  
Grace S. Leung ◽  
Ralph R. Isberg
Keyword(s):  
Dictyostelium Discoideum ◽  
Mycobacterium Marinum ◽  
Intracellular Replication ◽  
Mycobacterial Species ◽  
Phagocytic Cells ◽  
Bacterial Mutant ◽  
Mouse Macrophages ◽  
Efficient Replication ◽  
Mycobacterial Growth

ABSTRACT Mycobacterium marinum causes tuberculosis-like disease in fish and amphibians and has been used as a model mycobacterial species because of its rapid growth and less stringent containment requirements relative to other mycobacterial species. We demonstrate here that M. marinum grows within Dictyostelium discoideum cells, allowing the genetic analysis of host factors that may modulate the replication of mycobacterial species. Intracellular growth of M. marinum was shown to mimic the properties previously observed for growth within cultured phagocytes. A defined bacterial mutant defective for growth within phagocytic cells was shown to be similarly defective for growth within D. discoideum. To test the role of host coronin, which was previously hypothesized to positively modulate mycobacterial growth within mouse macrophages, a defined D. discoideum coronin mutant was analyzed. Surprisingly, the absence of coronin resulted in enhanced intracellular replication of M. marinum relative to the control wild-type strain. Consistent with previous observations, some phagosomes showed persistence of coronin about the surface of the compartment, but colocalization of the protein was far from uniform. We conclude that in D. discoideum factors other than coronin support intracellular replication of M. marinum.

scholarly journals Inhibitors of Mycobacterium marinum virulence identified in a Dictyostelium discoideum host model

PLoS ONE ◽  
2017 ◽  
Vol 12 (7) ◽  
pp. e0181121 ◽  
Author(s):  
Hajer Ouertatani-Sakouhi ◽  
Sébastien Kicka ◽  
Gianpaolo Chiriano ◽  
Christopher F. Harrison ◽  
Hubert Hilbi ◽  
...  
Keyword(s):  
Dictyostelium Discoideum ◽  
Mycobacterium Marinum

scholarly journals The Dictyostelium discoideum E3 ubiquitin ligase TrafE coordinates endolysosomal damage response and cell-autonomous immunity to Mycobacterium marinum.

2021 ◽  
Author(s):  
Lyudmil Raykov ◽  
Manon Mottet ◽  
Jahn Nitschke ◽  
Thierry Soldati
Keyword(s):  
Cell Death ◽  
Dictyostelium Discoideum ◽  
Cellular Response ◽  
Membrane Damage ◽  
Mycobacterium Marinum ◽  
Chemical Components ◽  
Key Factors ◽  
Endosomal Sorting ◽  
Damage Response ◽  
Intracellular Compartments

Cells are perpetually challenged by pathogens, protein aggregates or chemicals, that induce plasma membrane or endolysosomal compartments damage. Endolysosomal perforations are recognised as severe stress, however the mechanisms of the cellular response that ensure quality control, repair and endolysosomal homeostasis are just beginning to be unravelled. The endosomal sorting complex required for transport (ESCRT) and the autophagy machinery are recruited to damaged membranes to either repair or to remove membrane remnants. Crucial element of the endolysosomal damage response (ELDR) are factors that sense damage, paralleled by extensive tagging of the damaged organelles with signals, such as ubiquitin, required for the recruitment of ELDR components. Unattended membrane damage leads to leakage of harmful components including protons and reactive oxygen species that cause cell death. To explore ELDR key factors responsible for detection and marking of damaged compartments we use the professional phagocyte Dictyostelium discoideum. We found an evolutionary conserved E3-ligase TrafE that is robustly recruited to intracellular compartments disrupted after infection with Mycobacterium marinum or after sterile damage caused by chemical components. Importantly, we show that the absence of TrafE severely compromises the xenophagy restriction of bacteria as well as autophagy-mediated and ESCRT-mediated ELDR, resulting in early cell death.

scholarly journals Proteomic characterization of the Mycobacterium marinum-containing vacuole in Dictyostelium discoideum

2019 ◽  
Author(s):  
Aurélie Guého ◽  
Cristina Bosmani ◽  
Thierry Soldati
Keyword(s):  
Dictyostelium Discoideum ◽  
Model System ◽  
Mycobacterium Marinum ◽  
Close Relative ◽  
Phagosome Maturation ◽  
Fish Pathogen ◽  
Free Living ◽  
Free Living Amoeba ◽  
Professional Phagocyte

ABSTRACTMycobacterium tuberculosis, the causative agent of tuberculosis, is able to manipulate the phagosome compartment where it resides in order to establish a permissive replicative compartment called the Mycobacterium-containing vacuole (MCV). Mycobacterium marinum, a fish pathogen and a close relative of the tuberculosis group, is also able to infect the free-living amoeba and professional phagocyte Dictyostelium discoideum and to manipulate its phagosome maturation. By using this host/pathogen model system, we have established an innovative procedure to isolate MCVs. This procedure allowed us to isolate M. marinum-MCV at 1, 3 and 6 hours post infection to study the early M. marinum-MCV proteome. By using isobaric labelling and mass spectrometry, we quantitatively compared the proteomic composition of those MCVs isolated at different stages of the early infection phase to understand how M. marinum impacts on this compartment to divert it from the normal phagosomal pathway. Furthermore, we also compare the manipulated compartment M. marinum-MCV to non- or less manipulated compartments containing different mycobacteria strains: the non-pathogenic M. smegmatis, the avirulent M. marinum-L1D or the attenuated M. marinum-RD1.

scholarly journals Time-resolved RNA-seq profiling of the infection of Dictyostelium discoideum by Mycobacterium marinum reveals an integrated host response to damage and stress

2019 ◽  
Author(s):  
N. Hanna ◽  
F. Burdet ◽  
A. Melotti ◽  
C. Bosmani ◽  
S. Kicka ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dictyostelium Discoideum ◽  
Host Response ◽  
Mycobacterium Marinum ◽  
Infection Model ◽  
Multiple Drug ◽  
Late Time ◽  
Membrane Repair ◽  
Time Resolved ◽  
Recent Emergence

ABSTRACTTuberculosis remains the most pervasive infectious disease and the recent emergence of multiple drug-resistant strains emphasizes the need for more efficient drug treatments. The experimentally versatile Dictyostelium discoideum – Mycobacterium marinum infection model provides a powerful system to study mycobacteria pathogenicity and host response. In this study, a time-resolved transcriptomic analysis of the amoeba D. discoideum was performed to decipher the different host pathways impacted during infection. We investigated how D. discoideum fine-tunes its gene expression in response to M. marinum infection by assessing the transcriptomic profile covering the critical stages of entry, establishment of a permissive niche, proliferation and dissemination (1, 3, 6, 12, 24 and 48 hours post infection). Differential gene expression provided a fingerprint of the transcriptome of the host cell in the presence of mycobacteria, and helped identify specific markers and molecular signatures of infection. Enrichment pathway analysis showed that most of the Biological Processes (BP) of upregulated genes at early time point of infection hinted towards damage response and cellular defence, especially in specific pathways involved in membrane repair (ESCRT) and bacteria elimination (autophagy). Whereas at late time points of infection, BP related to starvation were upregulated. Some other signatures were more unexpected, such as cell cycle (downregulation of cytosolic large & small ribosomal subunits) and upregulation of metabolic adaptations (lipids transport).

scholarly journals Zn2+ intoxication of Mycobacterium marinum during Dictyostelium discoideum infection is counteracted by induction of the pathogen Zn2+ exporter CtpC

2019 ◽  
Author(s):  
Louise H. Lefrançois ◽  
Vera Kalinina ◽  
Elena Cardenal-Muñoz ◽  
Nabil Hanna ◽  
Hendrik Koliwer-Brandl ◽  
...  
Keyword(s):  
Dictyostelium Discoideum ◽  
Metal Accumulation ◽  
Chemical Warfare ◽  
Mycobacterium Marinum ◽  
Close Relative ◽  
First Time ◽  
Intracellular Proliferation ◽  
Role Of Zn ◽  
Professional Phagocyte

ABSTRACTMacrophages use diverse strategies to kill or restrict intracellular pathogens. Some of these strategies involve the deprivation of bacteria from (micro)nutrients such as transition metals, and the bacteria intoxication through metal accumulation. Little is known about the chemical warfare between Mycobacterium marinum, a close relative of the human pathogen M. tuberculosis, and its hosts. Here we use the professional phagocyte Dictyostelium discoideum to investigate the role of Zn2+ during M. marinum infection. We show that M. marinum infection induces the accumulation of Zn2+ inside the Mycobacterium-containing vacuole (MCV), achieved by the induction and recruitment of the D. discoideum Zn2+ efflux pumps ZntA and ZntB. In cells lacking the ZntA detoxifying transporter there is further attenuation of M. marinum growth, possibly due to a compensatory efflux of Zn2+ into the MCV. This efflux is presumably carried out by ZntB, the main Zn2+ transporter in endosomes and phagosomes. Counterintuitively, M. marinum growth is also impaired in zntB KO cells, where MCVs accumulate less Zn2+. We also demonstrate that M. marinum senses toxic levels of Zn2+ and responds by upregulating its Zn2+ exporter CtpC, which supports bacteria survival under these restrictive conditions. Attenuation of M. marinum intracellular proliferation in zntA and zntB KO cells is accentuated in the absence of CtpC, confirming that mycobacteria face noxious levels of Zn2+. Altogether, we show for the first time that M. marinum infection induces a deleterious Zn2+ elevation in D. discoideum, which is counteracted by the bacteria with the induction of its Zn2+ exporter CtpC.

scholarly journals ESCRT and autophagy cooperate to repair ESX-1-dependent damage to the Mycobacterium-containing vacuole

2018 ◽  
Author(s):  
Ana T. López-Jiménez ◽  
Elena Cardenal-Muñoz ◽  
Florence Leuba ◽  
Lilli Gerstenmaier ◽  
Monica Hagedorn ◽  
...  
Keyword(s):  
Dictyostelium Discoideum ◽  
Membrane Damage ◽  
Mycobacterium Marinum ◽  
Eukaryotic Cells ◽  
Intracellular Pathogens ◽  
Membrane Repair ◽  
Endosomal Sorting

AbstractPhagocytes capture invader microbes within the bactericidal phagosome. Some pathogens subvert killing by damaging and escaping from this compartment. To prevent and fight bacterial escape, cells contain and repair the membrane damage, or finally eliminate the cytosolic escapees. All eukaryotic cells engage highly conserved mechanisms to ensure integrity of membranes in a multitude of physiological and pathological situations, including the Endosomal Sorting Complex Required for Transport (ESCRT) and autophagy machineries. In Dictyostelium discoideum, recruitment of the ESCRT-III protein Snf7/Chmp4/Vps32 and the ATPase Vps4 to sites of membrane repair relies on the ESCRT-I component Tsg101 and occurs in absence of Ca2+. The ESX-1 dependent membrane perforations produced by the pathogen Mycobacterium marinum separately engage both ESCRT and autophagy. In absence of Tsg101, M. marinum escapes earlier to the cytosol, where it is restricted by xenophagy. We propose that ESCRT has an evolutionary conserved function in containing intracellular pathogens in intact compartments.

scholarly journals A Mycobacterium marinum TesA mutant defective for major cell wall-associated lipids is highly attenuated in Dictyostelium discoideum and zebrafish embryos

2011 ◽  
Vol 80 (4) ◽  
pp. 919-934 ◽  
Author(s):  
Laeticia Alibaud ◽  
Yoann Rombouts ◽  
Xavier Trivelli ◽  
Adeline Burguière ◽  
Suat L. G. Cirillo ◽  
...  
Keyword(s):  
Cell Wall ◽  
Dictyostelium Discoideum ◽  
Mycobacterium Marinum ◽  
Zebrafish Embryos
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