Mycobacterium tuberculosis Exploits Asparagine to Assimilate Nitrogen and Resist Acid Stress during Infection

Gouzy, Alexandre; Larrouy-Maumus, Gérald; Bottai, Daria; Levillain, Florence; Dumas, Alexia; Wallach, Joshua B.; Caire-Brändli, Irène; Chastellier, Chantal de; Wu, Ting-Di; Poincloux, Renaud; Brosch, Roland; Guerquin‐Kern, Jean‐Luc; Schnappinger, Dirk; Carvalho, Luiz Pedro S. de; Poquet, Yannick; Neyrolles, Olivier

doi:10.1371/journal.ppat.1003928

Cited by 143 publications

(135 citation statements)

References 67 publications

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“…Subverting nutritional restrictions by the host (9, 35) via an autarkic metabolic lifestyle is likely an evolutionary advantage that contributes greatly to Mtb's unmatched persistence in humans. It turns out that this important feature of Mtb's metabolism also is a substantial vulnerability, because several auxotrophic strains of Mtb show significant levels of attenuation in mice (8,16,17,26,(36)(37)(38)(39)(40)(41)(42). However, most of these auxotrophies are less bactericidal than Fig.…”

Section: Discussionmentioning

confidence: 99%

Essential roles of methionine and S -adenosylmethionine in the autarkic lifestyle of Mycobacterium tuberculosis

Berney

Berney-Meyer

Wong

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

119

163

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Multidrug resistance, strong side effects, and compliance problems in TB chemotherapy mandate new ways to kill Mycobacterium tuberculosis (Mtb). Here we show that deletion of the gene encoding homoserine transacetylase (metA) inactivates methionine and S-adenosylmethionine (SAM) biosynthesis in Mtb and renders this pathogen exquisitely sensitive to killing in immunocompetent or immunocompromised mice, leading to rapid clearance from host tissues. Mtb ΔmetA is unable to proliferate in primary human macrophages, and in vitro starvation leads to extraordinarily rapid killing with no appearance of suppressor mutants. Cell death of Mtb ΔmetA is faster than that of other auxotrophic mutants (i.e., tryptophan, pantothenate, leucine, biotin), suggesting a particularly potent mechanism of killing. Time-course metabolomics showed complete depletion of intracellular methionine and SAM. SAM depletion was consistent with a significant decrease in methylation at the DNA level (measured by single-molecule real-time sequencing) and with the induction of several essential methyltransferases involved in biotin and menaquinone biosynthesis, both of which are vital biological processes and validated targets of antimycobacterial drugs. Mtb ΔmetA could be partially rescued by biotin supplementation, confirming a multitarget cell death mechanism. The work presented here uncovers a previously unidentified vulnerability of Mtb-the incapacity to scavenge intermediates of SAM and methionine biosynthesis from the host. This vulnerability unveils an entirely new drug target space with the promise of rapid killing of the tubercle bacillus by a new mechanism of action.host-pathogen interaction | bactericidal auxotrophy | amino acid biosynthesis | metabolism U nderstanding the metabolic interactions between an invading microbe and its host is becoming a new cornerstone of host-pathogen research (1-3). Many intracellular pathogens modulate the host response to satisfy their nutritional needs and as a result have become auxotrophic for several essential amino acids and cofactors (4-6). Mycobacterium tuberculosis (Mtb), arguably the most deadly bacterial pathogen in the world (7), adopted a different strategy. This ultra-slow-growing bacterium is prototrophic for all essential cofactors and amino acids, suggesting that it either dwells in host compartments where such metabolites are unavailable or actively chooses this autarkic lifestyle to retain metabolic flexibility and remain invisible to the host. Indeed, much of Mtb's long-term success as a human pathogen is ascribed to its extraordinary stealth in the face of host immunity (8, 9); Mtb's ability to evade detection by the host might explain why devising an efficient vaccine has failed thus far and why drug therapy is difficult. Therefore, understanding Mtb's in vivo metabolic requirements could help in the development of much-needed new strategies for antimycobacterial therapy.Methionine and S-adenosylmethionine (SAM) are essential metabolites that have gained considerable scientific attenti...

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Section: Discussionmentioning

confidence: 99%

Essential roles of methionine and S -adenosylmethionine in the autarkic lifestyle of Mycobacterium tuberculosis

Berney

Berney-Meyer

Wong

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

119

163

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“…As a result, there have been wide-ranging suggestions about its actual function. Recently, it was reported that M. tuberculosis secretes an asparaginase, AnsA, to hydrolyze asparagine and generate ammonium ions, which can then block phagosomal acidification (62). This protein was shown to partially depend on the ESX-5 secretion system for export.…”

Section: Discussionmentioning

confidence: 99%

A Duplicated ESAT-6 Region of ESX-5 Is Involved in Protein Export and Virulence of Mycobacteria

et al. 2015

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The ESX-5 secretion system of Mycobacterium tuberculosis is important for bacterial virulence and for the secretion of the large PE/PPE protein family, whose genes constitute 10% of the M. tuberculosis genome. A four-gene region of the ESX-5 system is duplicated three times in the M. tuberculosis genome, but the functions of these duplicates are unknown. Here we investigated one of these duplicates: the region carrying the esxI, esxJ, ppe15, and pe8 genes (ESX-5a). An ESX-5a deletion mutant in the model system M. marinum background was deficient in the secretion of some members of the PE/PPE family of proteins. Surprisingly, we also identified other proteins that are not members of this family, thus expanding the range of ESX-5 secretion substrates. In addition, we demonstrated that ESX-5a is important for the virulence of M. marinum in the zebrafish model. Furthermore, we showed the role of the M. tuberculosis ESX-5a region in inflammasome activation but not host cell death induction, which is different from the case for the M. tuberculosis ESX-5 system. In conclusion, the ESX-5a region is nonredundant with its ESX-5 paralog and is necessary for secretion of a specific subset of proteins in M. tuberculosis and M. marinum that are important for bacterial virulence of M. marinum. Our findings point to a role for the three ESX-5 duplicate regions in the selection of substrates for secretion via ESX-5, and hence, they provide the basis for a refined model of the molecular mechanism of this type VII secretion system. Mycobacterium tuberculosis is an extremely successful pathogen that employs various strategies to evade immune responses and persist within the host (1, 2). Integral to this manipulation of the host by M. tuberculosis is the secretion of virulence factors by various secretion systems (3). There are five different type VII secretion systems (T7SS) in M. tuberculosis (ESX-1 to ESX-5) (4, 5). The different ESX systems were most likely generated via duplications of the ancestral system ESX-4 in the chronological order ESX-1, ESX-3, ESX-2, and ESX-5 (6). All the ESX secretion systems have a set of common genes which encode the core components of their secretion machinery, among which are genes encoding two members of the ESAT-6-like family (Esx proteins) (5, 7).The region of difference 1 (RD1) is a 9.5-kbp stretch comprising 9 genes of ESX-1 that is deleted in all strains of the live tuberculosis vaccine M. bovis BCG (8). The importance of ESX-1 for virulence was first demonstrated by expressing the entire RD1 locus in the BCG vaccine strain and restoring virulence in the mouse model of tuberculosis (9, 10). ESX-1 is essential for full virulence of M. tuberculosis in mice (11-13). In M. marinum, it is required for the growth of bacteria within macrophages, cell-tocell spread, and virulence in the zebrafish model (14). ESX-1 is required for the cosecretion of two ESAT-6 family proteins, EsxA and EsxB (13). The other known substrates are Rv3881c (15) and Rv3864 (16).The ESX-5 system is the most recently evolve...

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“…S. Typhimurium is a facultative intracellular pathogen that thrives inside phagocytes, where access to nutrients is limited. To determine if S. Typhimurium, like Mycobacterium tuberculosis (20) and Francisella tularensis (21), catabolizes asparagine to survive and replicate inside phagocytes, we infected bone marrow-derived macrophages cultured from 129X1/SvJ mice with wild-type S. Typhimurium, the ⌬STM1294 single mutant, the ⌬STM3106 single mutant, or the ⌬STM1294 ⌬STM3106 double mutant. After 18 h of infection, we recovered intracellular bacteria and found reduced numbers of the ⌬STM1294 mutant and the ⌬STM1294 ⌬STM3106 mutant relative to the level of the wild-type S. Typhimurium, while the ⌬STM3106 mutant followed the same trend ( Fig.…”

Section: Stm3997 Is Required For S Typhimurium To Cause Inhibition Omentioning

confidence: 99%

Contribution of Asparagine Catabolism to Salmonella Virulence

et al. 2017

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Salmonellae are pathogenic bacteria that cause significant morbidity and mortality in humans worldwide. Salmonellae establish infection and avoid clearance by the immune system by mechanisms that are not well understood. We previously showed that L-asparaginase II produced by Salmonella enterica serovar Typhimurium (S. Typhimurium) inhibits T cell responses and mediates virulence. In addition, we previously showed that asparagine deprivation such as that mediated by L-asparaginase II of S. Typhimurium causes suppression of activation-induced T cell metabolic reprogramming. Here, we report that STM3997, which encodes a homolog of disulfide bond protein A (dsbA) of Escherichia coli, is required for L-asparaginase II stability and function. Furthermore, we report that L-asparaginase II localizes primarily to the periplasm and acts together with L-asparaginase I to provide S. Typhimurium the ability to catabolize asparagine and assimilate nitrogen. Importantly, we determined that, in a murine model of infection, S. Typhimurium lacking both L-asparaginase I and II genes competes poorly with wild-type S. Typhimurium for colonization of target tissues. Collectively, these results indicate that asparagine catabolism contributes to S. Typhimurium virulence, providing new insights into the competition for nutrients at the host-pathogen interface.

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Mycobacterium tuberculosis Exploits Asparagine to Assimilate Nitrogen and Resist Acid Stress during Infection

Cited by 143 publications

References 67 publications

Essential roles of methionine and S -adenosylmethionine in the autarkic lifestyle of Mycobacterium tuberculosis

Essential roles of methionine and S -adenosylmethionine in the autarkic lifestyle of Mycobacterium tuberculosis

A Duplicated ESAT-6 Region of ESX-5 Is Involved in Protein Export and Virulence of Mycobacteria

Contribution of Asparagine Catabolism to Salmonella Virulence

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