AF Alice

AK Pandey

B Buschmeier

B Jayanthi

C Stover

CM Sassetti

D Fraenkel

D Schnappinger

EJ Muñoz-Elías

F Daldal

F Foufelle

G Paterson

HIM Boshoff

J Babul

J Marrero

J McKinney

JH Shin

K Pethe

KC Yam

Kevin Pethe

L Miallau

L Shi

LA Kelley

LG Wayne

LMD Orchard

LPS de Carvalho

M Schumacher

MI Voskuil

MR Hasan

MR Lovingshimer

PC Hsieh

R Dubos

R Kalscheuer

R Kapoor

S Aly

S Bardarov

SD Bowden

Srinivasa P. S. Rao

ST Cole

Sylvie Alonso

T Morita

T Parish

Tanya Parish

Thomas Dick

W Malaga

Wai Yee Phong

Wenwei Lin

PLoS ONE

PubMed

Crossref

Characterization of Phosphofructokinase Activity in Mycobacterium tuberculosis Reveals That a Functional Glycolytic Carbon Flow Is Necessary to Limit the Accumulation of Toxic Metabolic Intermediates under Hypoxia

Metabolic versatility has been increasingly recognized as a major virulence mechanism that enables Mycobacterium tuberculosis to persist in many microenvironments encountered in its host. Glucose is one of the most abundant carbon sources that is exploited by many pathogenic bacteria in the human host. M. tuberculosis has an intact glycolytic pathway that is highly conserved in all clinical isolates sequenced to date suggesting that glucose may represent a non-negligible source of carbon and energy for this pathogen in vivo. Fructose-6-phosphate phosphorylation represents the key-committing step in glycolysis and is catalyzed by a phosphofructokinase (PFK) activity. Two genes, pfkA and pfkB have been annotated to encode putative PFK in M. tuberculosis. Here, we show that PFKA is the sole PFK enzyme in M. tuberculosis with no functional redundancy with PFKB. PFKA is required for growth on glucose as sole carbon source. In co-metabolism experiments, we report that disruption of the glycolytic pathway at the PFK step results in intracellular accumulation of sugar-phosphates that correlated with significant impairment of the cell viability. Concomitantly, we found that the presence of glucose is highly toxic for the long-term survival of hypoxic non-replicating mycobacteria, suggesting that accumulation of glucose-derived toxic metabolites does occur in the absence of sustained aerobic respiration. The culture medium traditionally used to study the physiology of hypoxic mycobacteria is supplemented with glucose. In this medium, M. tuberculosis can survive for only 7-10 days in a true non-replicating state before death is observed. By omitting glucose in the medium this period could be extended for up to at least 40 days without significant viability loss. Therefore, our study suggests that glycolysis leads to accumulation of glucose-derived toxic metabolites that limits long-term survival of hypoxic mycobacteria. Such toxic effect is exacerbated when the glycolytic pathway is disrupted at the PKF step

Srinivasa P S Rao

Directory of Open Access Journals

Characterization of phosphofructokinase activity in Mycobacterium tuberculosis reveals that a functional glycolytic carbon flow is necessary to limit the accumulation of toxic metabolic intermediates under hypoxia.

Correction: Characterization of Phosphofructokinase Activity in Mycobacterium tuberculosis Reveals That a Functional Glycolytic Carbon Flow Is Necessary to Limit the Accumulation of Toxic Metabolic Intermediates under Hypoxia

Phong Wai Yee

Lin Wenwei

Rao Srinivasa P. S.

Dick Thomas

Alonso Sylvie

Pethe Kevin

Public Library of Science (PLOS)

 Reveals That a Functional Glycolytic Carbon Flow Is Necessary to Limit the Accumulation of Toxic Metabolic Intermediates under Hypoxia

Mycobacterium tuberculosis metabolic versatility has been increasingly recognized as a major virulence mechanism enabling this pathogen to persist in many microenvironments encountered in its host. Glucose is one of the most abundant source of carbon that is exploited by many pathogenic bacteria in the human host. M. tuberculosis has an intact glycolytic pathway that is highly conserved in all clinical isolates sequenced to date suggesting that, in addition to lipids, glucose may represent a non-negligible source of carbon and energy for this pathogen in vivo. Fructose-6-phosphate phosphorylation represents the key-committing step in glycolysis and is catalyzed by phosphofructokinase (PFK). Two genes, pfkA and pfkB, have been annotated to encode putative PFK in M. tuberculosis. Here, we show that PFKA is the sole PFK enzyme in M. tuberculosis with no functional redundancy with PFKB. PFKA is required for growth on glucose as sole carbon source. Furthermore, in co-metabolism experiments, a disrupted glycolytic pathway resulted in decreased survival due to the accumulation of glucose-derived toxic intermediate metabolites. Coincidentally we found that glucose metabolism is highly toxic for the long term survival of hypoxic non-replicating M. tuberculosis. Indeed, M. tuberculosis survived several order of magnitudes better in a glucose-depleted culture medium, compared to what is traditionally achieved in the original glucose-supplemented medium. This novel finding improves the potential and relevance of the Wayne model for the study of the mechanisms of persistence in M. tuberculosis. In conclusion, although a functional glycolytic pathway is not required for infection and persistence in the mouse model, we propose that glycolysis is required for regulating the pool of sugar phosphate that may be otherwise toxic for hypoxic mycobacteria

Phong, Wai Yee

Lin, WenWei

Rao, Srinivasa P.S.

Dick, Thomas

Sylvie, Alonso

Pethe, Kevin

The Novartis Repository

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World Health Organization

http://dx.doi.org/10.1371/journal.pone.0056037

Characterization of Phosphofructokinase Activity in Mycobacterium tuberculosis Reveals That a Functional Glycolytic Carbon Flow Is Necessary to Limit the Accumulation of Toxic Metabolic Intermediates under Hypoxia

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